Acylated aminopropanediols and analogues and therapeutic uses thereof

ABSTRACT

The invention relates to novel acylated aminopropanediols and the nitrogen and sulfur analogues thereof, pharmaceutical compositions comprising same, therapeutic uses thereof, in particular for the treatment of cerebral ischemia. The invention also provides a method of preparing said derivatives.

The invention relates to novel acylated aminopropanediols and thenitrogen and sulfur analogues thereof, pharmaceutical compositionscomprising same, therapeutic uses thereof, in particular for thetreatment of cerebral ischemia. The invention also provides a method ofpreparing said derivatives.

The inventive compounds have advantageous antioxidant andanti-inflammatory pharmacological properties. The invention alsodescribes methods of therapeutic treatment using said compounds andpharmaceutical compositions comprising same. In particular, theinventive compounds are useful for preventing or treating stroke.

In France, cerebrovascular disease (150,000 new cases annually) is thethird leading cause of mortality and the leading cause of disability inadults. Ischemic and hemorrhagic stroke respectively account for 80% and20% of all cerebrovascular accidents. Ischemic stroke is an importanttherapeutic issue that must be addressed in order to reduce themorbidity and mortality of cerebrovascular disease. Progress has beenmade not only in treating the acute phase of ischemia but also inpreventing same. It is therefore important to keep in mind that theidentification and management of risk factors are essential in thetreatment of this pathology.

Drug-based treatments of cerebral ischemia are based on differentstrategies. A first strategy comprises preventing the occurrence ofcerebral ischemic accidents through prevention of risk factors(hypertension, hypercholesterolemia, diabetes, atrial fibrillation,etc.) or through prevention of thrombosis, in particular with the helpof antiplatelet drugs or anticoagulants (Adams 2002) and (Gorelick2002).

A second strategy comprises treating the acute phase of ischemia, so asto attenuate its long-term consequences (Lutsep and Clark 2001).

The pathophysiology of cerebral ischemia can be described as follows:the ischemic penumbra, an intermediate zone between the ischemic focuswhere the neurons are necrotized and the intact nerve tissue, is thesite of a pathophysiological cascade which leads over the course of afew days to neuronal death, if reperfusion does not occur or ifneuroprotection is insufficient. The first event, which takes place inthe first few hours, is a massive release of glutamate which leads toneuron depolarization and cellular oedema. Calcium influx into the cellinduces mitochondrial damage leading to the release of free radicals andthe induction of enzymes that promote degradation of neuronal membranes.Calcium influx into the cell induces mitochondrial damage leading to therelease of free radicals and the induction of enzymes that promotedegradation of neuronal membranes. Calcium influx and free radicalproduction in turn activate certain transcription factors, such asNF-κB. Said activation induces inflammatory processes such as inductionof endothelial adhesion proteins, polynuclear neutrophil infiltration ofthe ischemic focus, microglial activation, induction of enzymes likenitric oxide (NO) synthase type II or cyclooxygenase type II. Theseinflammatory processes lead to release of NO or prostanoids which aretoxic to the cell. Together, these processes result in a phenomenon ofapoptosis inducing irreversible lesions (Dirnagl, ladecola et al. 1999).

The concept of prophylactic neuroprotection is based on experimentaldata in animal models demonstrating ischemic tolerance. In fact,different procedures applied prior to experimentally induced brainischemia attenuate the severity of the latter. Various stimuli caninduce brain ischemic tolerance:preconditioning (brief ischemiapreceding prolonged ischemia); heat stress; administration of a low doseof bacterial lipopolysaccharide (Bordet, Deplanque et al. 2000).

Said stimuli induce tolerance mechanisms which activate signalstriggering protective mechanisms. Different triggering mechanisms havebeen identified: cytokines, inflammatory pathways, free radicals, NO,ATP-dependent potassium channels, adenosine. The observed lag timebetween the onset of early events and ischemic tolerance stems from theneed for protein synthesis. Various types of proteins have been shown toinduce ischemic tolerance: heat shock proteins, antioxidant enzymes andanti-apoptotic proteins (Nandagopal, Dawson et al. 2001).

Thus there is a real need for compounds capable of preventing thedevelopment of risk factors for cerebrovascular accidents such asatherosclerosis, diabetes, obesity, and the like, capable of providingprophylactic neuroprotection but also active neuroprotection in theacute phase of cerebral ischemia.

The PPARs (α, β, γ) belong to the hormone-activated nuclear receptorfamily. When activated by binding with their ligand, they heterodimerizewith Retinoid-X-Receptor (RXR) and bind to “Peroxisome ProliferatorResponse Elements” (PPREs) located in the promoter sequence of targetgenes. Binding of PPAR to PPRE thereby induces expression of the targetgene (Fruchart, Staels et al. 2001).

The PPARs are distributed in a wide variety of organs, although they allexhibit a certain degree of tissue specificity with the exception ofPPARβ the expression of which appears to be ubiquitous. PPARα expressionis particularly high in liver and in the intestinal lumen whereas PPARγis expressed mainly in fat tissue and spleen. The three subtypes (α, β,γ) are expressed in the central nervous system. Cells such asoligodendrocytes and astrocytes more particularly express the PPARαsubtype (Kainu, Wikstrom et al. 1994).

The target genes of PPARs control lipid and glucose metabolism. However,recent discoveries suggest that the PPARs participate in otherbiological processes. PPAR activation by their ligands induces changesin the transcriptional activity of genes which modulate the inflammatoryprocess, antioxidant enzymes, angiogenesis, cell proliferation anddifferentiation, apoptosis, the activities of iNOS, MMPases and TIMPs(Smith, Dipreta et al. 2001) and (Clark 2002).

Free radicals play a role in a very wide range of pathologies includingallergy, tumor initiation and promotion, cardiovascular diseases(atherosclerosis, ischemia), genetic and metabolic disorders (diabetes),infectious and degenerative diseases (prion, etc.) and in ophthalmicdisorders (Mates, Perez-Gomez et al. 1999).

Reactive oxygen species (ROS) are produced during normal cellfunctioning. ROS comprise the hydroxyl radical (OH]), superoxide anion(O₂ ⁻), hydrogen peroxide (H₂O₂) and nitric oxide (NO). Said species arevery labile and, due to their high chemical reactivity, constitute adanger to the biological functions of cells. They induce lipidperoxidation, oxidation of certain enzymes and very extensive oxidationof proteins leading to degradation thereof. Protection against lipidperoxidation is a vital process in aerobic organisms, becauseperoxidation products can cause DNA damage. Thus a deregulation ormodification of the equilibrium between the production, processing andelimination of radical species by natural antioxidant defenses leads tothe establishment of processes that are deleterious to the cell ororganism.

ROS are processed via an antioxidant system that comprises an enzymaticcomponent and a non-enzymatic component. The enzymatic system iscomposed of several enzymes which have the following characteristics:

-   -   Superoxide dismutase (SOD) destroys the superoxide radical by        converting it to peroxide. The peroxide in turn is acted upon by        another enzyme system. Low levels of SOD are continuously        produced by aerobic respiration. Three classes of SOD have been        identified in humans, each containing Cu, Zn, Fe, Mn, or Ni as        cofactor. The three forms of human SOD are distributed as        follows: a cytosolic Cu-Zn SOD, a mitochondrial Mn—SO and an        extracellular SOD.    -   Catalase is very efficient at converting hydrogen peroxide        (H₂O₂) to water and oxygen. Hydrogen peroxide is enzymatically        catabolized in aerobic organisms. Catalase also catalyzes the        reduction of a variety of hydroperoxides (ROOH).    -   Glutathione peroxidase uses selenium as cofactor and catalyzes        the reduction of hydroperoxides (ROOH and H₂O₂) by using        glutathione, and thereby protects cells against oxidative        damage.

Non-enzymatic antioxidant defenses of cells comprise molecules which aresynthesized or supplied in the diet.

Antioxidant molecules are present in different cell compartments.Detoxification enzymes for example eliminate free radicals and areessential to cell life. The three most important types of antioxidantcompounds are the carotenoids, vitamin C and vitamin E (Gilgun-Sherki,Melamed et al. 2001).

To avoid the phenomenon of apoptosis induced by cerebral ischemia andits resultant effects, the inventors have developed novel compoundscapable of preventing the development of the risk factors describedearlier and capable of exerting a prophylactic neuroprotective activity,but also of providing active neuroprotection during the acute phase ofcerebral ischemia.

The inventors have also shown that the compounds according to theinvention concurrently display PPAR activator, antioxidant andanti-inflammatory properties and, as such, said compounds have animportant therapeutic or prophylactic potential in cerebral ischemia.

The present invention thus provides a novel family of compoundsexhibiting advantageous pharmacological properties useful for thepreventive or curative treatment of cerebral ischemia. The inventionalso provides for methods for preparing said derivatives.

The compounds of the invention are represented by general formula (I):

in which:

-   -   G2 and G3 independently represent an oxygen atom, a sulfur atom        or N-R4 group, G2 and G3 not simultaneously representing a N-R4        group,    -   R and R4 independently represent a hydrogen atom or a linear or        branched alkyl group, saturated or not, optionally substituted,        containing from 1 to 5 carbon atoms,    -   R1, R2 and R3, which are the same or different, represent a        hydrogen atom, a CO-R5 group or a group corresponding to the        formula CO—(CH₂)_(2n+1)—X—R6, at least one of the groups R1, R2        or R3 being a group corresponding to the formula        CO—(CH₂)_(2n+1)—X—R6,    -   R5 is a linear or branched alkyl group, saturated or not,        optionally substituted, possibly comprising a cyclic group, the        main chain of which contains from 1 to 25 carbon atoms,    -   X is a sulfur atom, a selenium atom, a SO group or a SO₂ group,    -   n is a whole number comprised between 0 and 11,    -   R6 is a linear or branched alkyl group, saturated or not,        optionally substituted, possibly comprising a cyclic group, the        main chain of which contains from 3 to 23 carbon atoms,        preferably 10 to 23 carbon atoms and optionally one or more        heterogroups selected in the group consisting of an oxygen atom,        a sulfur atom, a selenium atom, a SO group and SO₂ group,    -   with the exception of compounds having formula (I) in which G2R2        and G3R3 simultaneously represent hydroxyl groups.

In compounds represented by general formula (I) according to theinvention, the R5 group or groups, which are the same or different,preferably represent a linear or branched alkyl group, saturated orunsaturated, substituted or not, the main chain of which contains from 1to 20 carbon atoms, even more preferably 7 to 17 carbon atoms, stillmore preferably 14 to 17. In compounds represented by general formula(I) according to the invention, the R5 group or groups, which are thesame or different, can also represent a lower alkyl group containing 1to 6 carbon atoms, such as in particular the methyl, ethyl, propyl,isopropyl, butyl, isobutyl, pentyl or hexyl group.

In compounds represented by general formula (I) according to theinvention, the R6 group or groups, which are the same or different,preferably represent a linear or branched alkyl group, saturated orunsaturated, substituted or not, the main chain of which contains from 3to 23 carbon atoms, preferably 13 to 20 carbon atoms, even morepreferably 14 to 17 carbon atoms, and still more preferably 14 carbonatoms.

Specific examples of saturated long chain alkyl groups for R5 or R6 arein particular the groups C₇H₁₅, C₁₀H₂₁, C₁₁H₂₃, C₁₃H₂₇, C₁₄H₂₉, C₁₅H₃₁,C₁₆H₃₃, C₁₇H₃₅. Specific examples of unsaturated long chain alkyl groupsfor R5 or R6 are in particular the groups C₁₄H₂₇, C₁₄H₂₅, C₁₅H₂₉,C₁₇H₂₉, C₁₇H₃₁, C₁₇H₃₃, C₁₉H₂₉, C₁₉H₃₁, C₂₁H₃₁, C₂₁H₃₅, C₂₁H₃₇, C₂₁H₃₉,C₂₃H₄₅ or the alkyl chains of eicosapentanoic (EPA) C_(20:5) (5, 8, 11,14, 17) and docosahexanoic (DHA) C_(22:6) (4, 7, 10, 13, 16, 19) acids.

Examples of branched long chain alkyl groups are in particular thegroups (CH₂)_(n), —CH(CH₃)C₂H₅ (CH═C(CH₃)—(CH₂)₂)_(n″)—CH═C(CH₃)₂ or(CH₂)_(2x+1)—C(CH₃)₂—(CH₂)_(n), —CH₃ (x being a whole number equal to orcomprised between 1 and 11, n′ being a whole number equal to orcomprised between 1 and 22, n″ being a whole number equal to orcomprised between 1 and 5, n′″ being a whole number equal to orcomprised between 0 and 22, and (2x+n′″) being less than or equal to 22,preferably less than or equal to 20).

As indicated earlier, the alkyl groups R5 or R6 can optionally comprisea cyclic group. Examples of cyclic groups are in particular cyclopropyl,cyclobutyl, cyclopentyl and cyclohexyl.

As indicated earlier, the alkyl groups R5 or R6 can optionally besubstituted by one or more substituents, which are the same ordifferent. The substituents are preferably selected in the groupconsisting of a halogen atom (iodine, chlorine, fluorine, bromine) and a—OH, ═O, —NO₂, —NH₂, —CN, —O—CH₃, —CH₂—OH, —CH₂OCH₃, —CF₃ and —COOZgroup (Z being a hydrogen atom or an alkyl group, preferably containingfrom 1 to 5 carbon atoms).

The invention also concerns the optical and geometrical isomers of saidcompounds, the racemates, salts, hydrates thereof and the mixturesthereof.

Compounds represented by formula (Ia) are compounds corresponding toformula (I) according to the invention in which a single one of thegroups R1, R2 or R3 represents a hydrogen atom.

Compounds represented by formula (Ib) are compounds corresponding toformula (I) according to the invention in which two of the groups R1, R2or R3 represent a hydrogen atom.

The invention also encompasses the prodrugs of the compounds representedby formula (I) which, after administration to a subject, are convertedto compounds represented by formula (I) and/or metabolites of compoundsrepresented by formula (I) which display therapeutic activities,particularly for the treatment of cerebral ischemia, which are similarto compounds represented by formula (I).

Moreover, in the group CO—(CH₂)_(2n+1)—X—R6, X most preferablyrepresents a sulfur or selenium atom and advantageously a sulfur atom.

Moreover, in the group CO—(CH₂)_(2n+1)—X—R6, n is preferably comprisedbetween 0 and 3, more specifically comprised between 0 and 2 and inparticular is equal to 0.

In the compounds represented by general formula (I) according to theinvention, R6 can contain one or more heterogroups, preferably 0, 1 or2, more preferably 0 or 1, selected in the group consisting of an oxygenatom, a sulfur atom, a selenium atom, a SO group or a SO₂ group.

A specific example of a CO—(CH₂)_(2n+1)—X—R6 group according to theinvention is the group CO—CH₂—S—C₁₄H₂₉. Preferred compounds in thespirit of the invention are therefore compounds represented by generalformula (I) hereinabove in which at least one of the groups R1, R2 andR3 represents a CO—(CH₂)_(2n+1)—X—R6 group in which X represents asulfur or selenium atom and preferably a sulfur atom and/or R6 is asaturated and linear alkyl group containing from 3 to 23 carbon atoms,preferably 13 to 20 carbon atoms, preferably 14 to 17, more preferably14 to 16, and even more preferably 14 carbon atoms.

Other particular compounds of the invention are those in which at leasttwo of the groups R1, R2 and R3 are CO—(CH₂)_(2n+1)—X—R6 groups, whichare the same or different, in which X represents a sulfur or seleniumatom and preferably a sulfur atom.

Particular compounds according to the invention are those in which G2represents an oxygen or sulfur atom, and preferably an oxygen atom. Insaid compounds, R2 advantageously represents a group corresponding tothe formula CO—(CH₂)_(2n+1)—X—R6 such as defined hereinabove.

Particularly preferred compounds are compounds represented by generalformula (I) hereinabove in which:

-   -   G3 is a N—R4 group in which R4 is a hydrogen atom or a methyl        group, and G2 is an oxygen atom; and/or    -   R2 represents a CO—(CH₂)_(2n+1)—X—R6 group such as defined        hereinabove.

Other preferred compounds are compounds represented by general formula(I) hereinabove in which R1, R2 and R3, which are the same or different,preferably the same, represent a CO—(CH₂)_(2n+1)—X—R6 group such asdefined hereinabove, in which X represents a sulfur or selenium atom andpreferably a sulfur atom and/or R6 represents a saturated and linearalkyl group containing from 13 to 17 carbon atoms, preferably 14 to 17,even more preferably 14 carbon atoms, in which n is preferably comprisedbetween 0 and 3, and in particular is equal to 0. More specifically,preferred compounds are compounds represented by general formula (I) inwhich R1, R2 and R3 represent CO—CH₂—S—C₁₄H₂₉ groups.

Examples of preferred inventive compounds are given in FIG. 1.

Thus, the invention more particularly has as object the compoundsrepresented by formula (I) selected from among:

-   1-tetradecylthioacetylamino-2,3-(dipalmitoyloxy)propane;-   3-tetradecylthioacetylamino-1,2-(ditetradecylthioacetyloxy)propane;-   3-palmitoylamino-1,2-(ditetradecylthioacetyloxy)propane;-   1,3-di(tetradecylthioacetylamino)propan-2-ol;-   1,3-diamino-2-(tetradecylthioacetyloxy)propane;-   1,3-ditetradecylthioacetylamino-2-(tetradecylthioacetyloxy)propane;-   1,3-dioleylamino-2-(tetradecylthioacetyloxy)propane;-   1,3-ditetradecylthioacetylamino-2-(tetradecylthioacetylthio)propane;    and-   1-tetradecylthioacetylamino-2,3-di(tetradecylthioacetylthio)propane.

The invention also has as object a pharmaceutical compositioncomprising, in a pharmaceutically acceptable support, at least onecompound represented by general formula (I) such as describedhereinabove, including compounds having formula (I) in which the groupsG2R2 and G3R3 simultaneously represent hydroxyl groups, possibly inassociation with another therapeutic agent. Said composition is intendedin particular to treat a cerebrovascular disease, such as cerebralischemia or hemorrhagic stroke.

Another object of the invention thus concerns any pharmaceuticalcomposition comprising in a pharmaceutically acceptable support at leastone compound represented by formula (I) such as described hereinabove,including compounds having formula (1) in which the groups G2R2 and G3R3simultaneously represent hydroxyl groups.

Advantageously it is a pharmaceutical composition for the treatment orprophylaxis of cerebrovascular pathologies and more particularlycerebral ischemia or cerebrovascular accidents. In fact, it was found ina surprising manner that compounds represented by formula (I), includingcompounds having formula (I) in which the groups G2R2 and G3R3simultaneously represent hydroxyl groups, concurrently display PPARactivator, antioxidant and anti-inflammatory properties and exhibitprophylactic and curative neuroprotective activity in cerebral ischemia.

The invention also concerns the use of a compound such as definedhereinabove for preparing a pharmaceutical composition intended forimplementing a method of treatment or prophylaxis in humans or animals.

The invention further concerns a method for treating cerebrovascularpathologies and more particularly cerebral ischemia, comprisingadministering to a subject, in particular human, an effective dose of acompound represented by formula (I) or of a pharmaceutical compositionsuch as defined hereinabove, including compounds having general formula(I) in which the groups G2R2 and G3R3 simultaneously represent hydroxylgroups.

Avantageously, the compounds represented by formula (I) which are usedare such as defined hereinabove and also comprise3-(tetradecylthioacetylamino)propane-1,2-diol.

The pharmaceutical compositions according to the inventionadvantageously comprise one or more pharmaceutically acceptibleexcipients or vehicles. Examples include pharmaceutically compatiblesaline, physiologic, isotonic, buffered solutions and the like, known tothose skilled in the art. The compositions may contain one or moreagents or vehicles selected from among dispersives, solubilizers,stabilizers, surfactants, preservatives, and the like. Agents orvehicles that may be used in the formulations (liquid and/or injectableand/or solid) comprise in particular methylcellulose,hydroxymethylcellulose, carboxymethylcellulose, polysorbate 80,mannitol, gelatin, lactose, vegetable oils, acacia and the like. Thecompositions may be formulated as injectable suspensions, gels, oils,tablets, suppositories, powders, gelatin capsules, capsules, and thelike, possibly by means of pharmaceutical forms or devices allowingsustained and/or delayed release. For this type of formulation, an agentsuch as cellulose, carbonates or starches is advantageously used.

The compounds or compositions of the invention may be administered indifferent ways and in different forms. For instance, they may beadministered systemically, by the oral route, parentally, by inhalationor by injection, such as for example by the intravenous, intramuscular,subcutaneous, transdermal, intra-arterial route, etc. For injections,the compounds are generally prepared in the form of liquid suspensions,which may be injected through syringes or by infusion, for instance. Inthis respect, the compounds are generally dissolved in pharmaceuticallycompatible saline, physiologic, isotonic, buffered solutions and thelike, known to those skilled in the art. For instance, the compositionsmay contain one or more agents or vehicles selected from amongdispersives, solubilizers, emulsifiers, stabilizers, surfactants,preservatives, buffers, and the like. Agents or vehicles that may beused in the liquid and/or injectable formulations comprise in particularmethylcellulose, hydroxymethylcellulose, carboxymethylcellulose,polysorbate 80, mannitol, gelatin, lactose, vegetable oils, acacia,liposomes, and the like.

The compositions may thus be administered in the form of gels, oils,tablets, suppositories, powders, gelatin capsules, capsules, aerosols,and the like, possibly by means of pharmaceutical forms or devicesallowing sustained and/or delayed release. For this type of formulation,an agent such as cellulose, carbonates or starches is advantageouslyused.

The compounds may be administered orally in which case the agents orvehicles used are preferably selected in the group consisting of water,gelatin, gums, lactose, starch, magnesium stearate, talc, an oil,polyalkylene glycol, and the like.

For parenteral administration, the compounds are preferably administeredin the form of solutions, suspensions or emulsions in particular withwater, oil or polyalkylene glycols to which, in addition topreservatives, stabilizers, emulsifiers, etc., it is also possible toadd salts to adjust osmotic pressure, buffers, and the like.

It is understood that the injection rate and/or injected dose may beadapted by those skilled in the art according to the patient, thepathology, the mode of administration, etc. Typically, the compounds areadministered at doses ranging from 1 μg to 2 g per dose, preferably from0.1 mg to 1 g per dose. The doses may be administered once a day orseveral times a day, as the case may be. Moreover, the compositions ofthe invention may also comprise other active substances or agents.

The invention also concerns methods for preparing the hereinabovecompounds. The compounds of the invention can be prepared fromcommercially available products, by employing a combination of chemicalreactions known to those skilled in the art.

According to one method of the invention, compounds represented byformula (I) in which (i) G2 and G3 are oxygen or sulfur atoms or a N—R4group, (ii) R and, as the case may be, R4, represent an identical linearor branched alkyl group, saturated or not, optionally substituted,containing from 1 to 5 carbon atoms and (iii) R1, R2 and R3, which arethe same or different, represent a CO—R5 group or a CO—(CH₂)_(2n+1)—X—R6group, are obtained from a compound represented by formula (I) in which(i) G2 or G3 are oxygen or sulfur atoms or a NH group, (ii) R is ahydrogen atom and (iii) R1, R2 and R3, which are the same or different,represent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group, an a compoundcorresponding to the formula A1-LG in which A1 represents the group Ror, as the case may be, R4 and LG is a reactive group selected forexample in the group consisting of Cl, Br, mesyl, tosyl, etc., possiblyin the presence of coupling agents or activators known to those skilledin the art.

In a first embodiment, compounds represented by formula (I) in which (i)G2 and G3 are oxygen or sulfur atoms or a NH group, (ii) R is a hydrogenatom and (iii) R1, R2 and R3, which are the same, represent aCO—(CH₂)_(2n+1)—X—R6 group, are obtained from a compound represented byformula (I) in which (i) G2 or G3 are oxygen or sulfur atoms or a NHgroup, (ii) R is a hydrogen atom and (iii) R1, R2 and R3 are hydrogenatoms and a compound corresponding to the formula A°-CO-A in which A isa reactive group selected for example in the group consisting of OH, Cl,O—CO-A° and O—R7, R7 being an alkyl group, and A° is the(CH₂)_(2n+1)—X—R6 group, possibly in the presence of coupling agents oractivators known to those skilled in the art.

Compounds represented by formula (I) according to the invention in which(i) G2 and G3 are oxygen atoms or a NH group, (ii) R is a hydrogen atomand (iii) R1, R2 and R3 are hydrogen atoms or represent a CO—R5 orCO—(CH₂)_(2n+1)—X—R6 group can be obtained by different methods whichenable the synthesis of compounds in which the groups carried on a sameheteroatom (nitrogen or oxygen) have the same meaning.

According to a first embodiment, a molecule of 1-aminoglycerol,1,3-diaminoglycerol or 1,2-diaminoglycerol (obtained by adapting theprotocol described by (Morris, Atassi et al. 1997)) is reacted with acompound corresponding to the formula A°-CO-A1 in which A1 is a reactivegroup selected for example in the group consisting of OH, Cl and OR7, R7being an alkyl group, and A° is the R5 group or the (CH₂)_(2n+1)—X—R6,group possibly in the presence of coupling agents or activators known tothose skilled in the art. Said reaction respectively yields particularforms of compounds represented by formula (I), named compounds (IIa-c),and can be carried out by adapting the protocols described by (Urakamiand Kakeda 1953), (Shealy, Frye et al. 1984), (Marx, Piantadosi et al.1988) and (Rahman, Ziering et al. 1988) or (Nazih, Cordier et al. 1999).In compounds (IIb-c), the groups carried on a same heteroatom,respectively, (R1 and R3) and (R1 and R2) have the same meaning.

Compounds represented by formula (I) according to the invention in which(i) G2 and G3 are oxygen atoms or a NH group, (ii) R is a hydrogen atomand (iii) R1, R2 and R3, which are the same or different, represent aCO—R5 or CO—(CH₂)_(2n+1)—X—R6 group, can be obtained from a compoundhaving formula (IIa-c) and a compound corresponding to the formulaA°-CO-A2 in which A2 is a reactive group selected for example in thegroup consisting of OH and Cl, and A° is the R5 group or the(CH₂)_(2n+1)—X—R6 group, possibly in the presence of coupling agents oractivators known to those skilled in the art. Said reaction enables thesynthesis of compounds in which the groups carried on a same heteroatom(nitrogen or oxygen), respectively (R1 and R2), (R1 and R3) or (R2 andR3) have the same meaning. Advantageously, said reaction is carried outaccording to the protocol described for example in (Urakami and Kakeda1953) and (Nazih, Cordier et al. 1999).

According to another particular method of the invention (diagram 1),compounds represented by formula (I) in which (i) G2 and G3 are oxygenatoms or a NH group (ii) R is a hydrogen atom and (iii) R1, R2 and R3,which are the same or different, represent a CO—R5 orCO—(CH₂)_(2n+1)—X—R5 group, can be obtained according to the followingsteps:

-   -   a) reacting 1-aminoglycerol, 1,3-diaminoglycerol or        1,2-diaminoglycerol with a compound (PG)₂O in which PG is a        protective group to give a compound having general formula        (IIIa-c). Advantageously, the reaction can be carried out by        adapting the protocols described by (Nazih, Cordier et al. 2000)        and (Kotsovolou, Chiou et al. 2001) in which (PG)₂O represents        di-tert-butyl dicarbonate;    -   b) reacting the compound having formula (IIIa-c) with a compound        corresponding to the formula A°-CO-A2 in which A2 is a reactive        group selected for example in the group consisting of OH and Cl,        and A° is the R5 group or the (CH₂)_(2n+1)—X—R6 group, possibly        in the presence of coupling agents or activators known to those        skilled in the art to give a compound represented by general        formula (IVa-c), in which R2 and R3 represent a CO—R5 or        CO—(CH₂)_(2n+1)—X—R6 group and PG is a protective group;    -   c) deprotecting the compound (IVa-c), according to conventional        conditions known to those skilled in the art, to give a compound        represented by general formula (I) in which (i) G2 and G3        represent an oxygen atom or a NH group, (ii) R and R1 are        hydrogen atoms and (iii) R2 and R3 represent a CO—R5 or        CO—(CH₂)_(2n+1)—X—R6 group;    -   d) reacting a compound represented by general formula (I) in        which (i) G2 and G3 represent an oxygen atom or a NH group, (ii)        R and R1 are hydrogen atoms and (iii) R2 and R3 represent a        CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group with a compound        corresponding to the formula A°-CO-A2 in which A2 is a reactive        group selected for example in the group consisting of OH and Cl,        and A° is the R5 group or the (CH₂)_(2n+1)—X—R6 group, possibly        in the presence of coupling agents or activators known to those        skilled in the art.

Compounds represented by formula (I) according to the invention in which(i) G2 and G3 are oxygen atoms, (ii) R is a hydrogen atom and (iii) R1,R2 and R3, which are the same or different, represent a CO—R5 orCO—(CH₂)_(2n+1)—X—R6 group, can be obtained in different ways.

According to a first method, a compound represented by formula (I)according to the invention, in which (i) G2 and G3 are oxygen atoms,(ii) R and R2 are hydrogen atoms and (iii) R1, R3, which are the same ordifferent, represent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group, is reactedwith a compound corresponding to the formula A°-CO-A2 in which A2 areactive group selected for example in the group consisting of OH andCl, and A° is the R5 group or the (CH₂)_(2n+1)—X—R6 group, possibly inthe presence of coupling agents or activators known to those skilled inthe art.

According to this method of preparation, compounds represented byformula (I) in which (i) G2 and G3 are oxygen atoms, (ii) R and R2 arehydrogen atoms and (iii) R1 and R3, which are the same or different,represent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group, can be obtained from acompound represented by formula (IIa) such as defined hereinabove and acompound corresponding to the formula A°-CO-A2 in which A2 is a reactivegroup selected for example in the group consisting of OH and Cl, and A°is the R5 group or the (CH₂)_(2n+1)—X—R6 group, possibly in the presenceof coupling agents or activators known to those skilled in the art.

According to another particular inventive method, compounds representedby formula (I) in which (i) G2 and G3 are oxygen atoms, (ii) R is ahydrogen atom and (iii) R1, R2 and R3, which are the same or different,represent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group, can be obtained from acompound represented by formula (I) according to the invention in which(i) G2 and G3 are oxygen atoms, (ii) R, R2 and R3 represent a hydrogenatom and (iii) R1 is a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group (compound offormula (IIa)) according to the following steps (diagram 2):

-   -   a) reacting a compound represented by formula (IIa) with a        compound PG-E in which PG is a protective group and E is a        reactive group selected for example in the group consisting of        OH and a halogen, to give a compound represented by general        formula (V) in which R1 is a CO—R5 or CO—(CH₂)_(2n+1)—X—R6        group. Advantageously, the reaction can be carried out by        adapting the protocols described by (Marx, Piantadosi et        al. 1988) and (Gaffney and Reese 1997) in which PG-E can        represent triphenylmethyl chloride or 9-phenylxanthene-9-ol or        else 9-chloro-9-phenylxanthene;    -   b) reacting a compound represented by formula (V) with a        compound corresponding to the formula A°-CO-A2 in which A2 is a        reactive group selected for example in the group consisting of        OH and Cl, and A° is the R5 group or the (CH₂)_(2n+1)—X—R6        group, possibly in the presence of coupling agents or activators        known to those skilled in the art to give a compound represented        by general formula (VI), in which R1 and R2, which are the same        or different, represent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group        and PG is a protective group;    -   c) deprotecting the compound (VI), in conditions known to those        skilled in the art, to give a compound represented by general        formula (I) in which (i) G2 and G3 are oxygen atoms, (ii) R and        R3 are hydrogen atoms and (iii) R1 and R2, which are the same or        different, represent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group;    -   d) reacting a compound represented by general formula (I) in        which (i) G2 and G3 are oxygen atoms, (ii) R and R3 are hydrogen        atoms and (iii) R1 and R2, which are the same or different,        represent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group with a compound        corresponding to the formula A°-C0-A2 in which A2 is a reactive        group selected for example in the group consisting of OH and Cl,        and A° is the R5 group or the (CH₂)_(2n+1)—X—R6 group, possibly        in the presence of coupling agents or activators known to those        skilled in the art.

In an advantageous manner, the hereinabove steps are carried outaccording to the protocols described by (Marx, Piantadosi et al. 1988).

According to another method of the invention, compounds represented byformula (I) in which (i) G2 or G3 represent an oxygen atom or a N—R4group, (ii) at least one of the groups G2 or G3 represents a N—R4 group,(iii) R and R4 independently represent linear or branched alkyl groups,saturated or not, optionally substituted, containing from 1 to 5 carbonatoms and (iv) R1, R2 and R3, which are the same or different, representa CO—R5 group or a CO—(CH₂)_(2n+1)—X—R6 group, are obtained by reactinga compound represented by formula (I) in which (i) one of the groupsG2R2 or G3R3 represents a hydroxyl group and the other group G2R2 orG3R3 represents a NR4R2 or NR4R3 group, respectively, with R2 or R3representing a CO—R5 group or a CO—(CH₂)_(2n+1)—X—R6 group, (ii) R andR4 independently represent a linear or branched alkyl group, saturatedor not, optionally substituted, containing from 1 to 5 carbon atoms and(iii) R1 represents a CO—R5 group or a CO—(CH2)_(2n+1)—X—R6 group, witha compound corresponding to the formula A°-CO-A2 in which A2 is areactive group selected for example in the group consisting of OH andCl, and A° is the R5 group or the (CH₂)_(2n+1)—X—R6 group, possibly inthe presence of coupling agents or activators known to those skilled inthe art.

Compounds represented by formula (I) according to the invention in which(i) one of the groups G2R2 or G3R3 represents a hydroxyl group and theother group G2R2 or G3R3 represents a NR4R2 or NR4R3 group,respectively, with R2 or R3 representing a CO—R5 group or aCO—(CH₂)_(2n+1)—X—R6 group, (ii) R and R4 independently represent linearor branched alkyl groups, saturated or not, optionally substituted,containing from 1 to 5 carbon atoms and (iii) R1 represents a CO—R5group or a CO—(CH₂)_(2n+1)—X—R6 group, are obtained from compoundsrepresented by formula (I) according to the invention in which one ofthe groups G2R2 or G3R3 represents a hydroxyl group and the other groupG2R2 or G3R3 represents a NR4R2 or NR4R3 group, respectively, with R2 orR3 representing a CO—R5 group or a CO—(CH₂)_(2n+1)—X—R6 group, (ii) Rand R4 independently represent a group such as defined hereinabove and(iii) R1 is a hydrogen atom and a compound corresponding to the formulaA°-C0-A2 in which A2 is a reactive group selected for example in thegroup consisting of OH and Cl, and A° is the R5 group or the(CH₂)_(2n+1)—X—R6 group, possibly in the presence of coupling agents oractivators known to those skilled in the art.

In a first embodiment, compounds represented by formula (I) according tothe invention in which (i) G2 is an oxygen atom, (ii) G3 represents aN—R4 group, (iii) R and R4 independently represent linear or branchedalkyl groups, saturated or not, optionally substituted, containing from1 to 5 carbon atoms, (iv) R1 and R2 are hydrogen atoms and (v) R3represents a CO—R5 group or a CO—(CH₂)_(2n+1)—X—R6 group are obtained inthe following manner (diagram 3):

-   -   a) reacting 1-aminoglycerol with a compound corresponding to the        formula R-CHO in which R represents a linear or branched alkyl        group, saturated or not, optionally substituted, containing from        1 to 5 carbon atoms and CHO is the aldehyde function in the        presence of reducing agents known to those skilled in the art to        give a compound represented by formula (VII) in which R is a        group such as defined hereinabove. Advantageously, said reaction        can be carried out by adapting the protocols described by        (Antoniadou-Vyzas, Foscolos et al. 1986);    -   b) reacting a compound represented by formula (VII) with a        compound (PG)₂O in which PG is a protective group to give a        compound represented by general formula (VIII). Advantageously,        the reaction can be carried out by adapting the protocols        described by (Nazih, Cordier et al. 2000) and (Kotsovolou, Chiou        et al. 2001) in which (PG)₂O represents di-tert-butyl        dicarbonate;    -   c) reacting a compound represented by formula (VIII) with a        compound corresponding to the formula LG-E in which E represents        a halogen and LG is a reactive group selected for example in the        group consisting of mesyl, tosyl, etc., to give a compound        represented by general formula (IX) by adapting the method        described by [Kitchin, Bethell et al. 1994];    -   d) reacting a compound represented by formula (IX) with a        compound corresponding to the formula R4-NH₂ in which R4        represents a linear or branched alkyl group, saturated or not,        optionally substituted, containing from 1 to 5 carbon atoms and        NH₂ represents the amine function, according to the method        described by (Ramalingan, Raju et al. 1995), to give a compound        corresonding to formula (X) in which R and R4, optionally        different, are such as defined hereinabove;    -   e) reacting a compound represented by formula (X) with a        compound corresponding to the formula A°-CO-A2 in which A2 is a        reactive group selected for example in the group consisting of        OH and Cl, and A° is the R5 group or the (CH₂)_(2n+1)—X—R6        group, possibly in the presence of coupling agents or activators        known to those skilled in the art to give a compound represented        by formula (XI) in which R and R4 represent linear or branched        alkyl groups, saturated or not, optionally substituted,        containing from 1 to 5 carbon atoms, R3 represents the R5 group        or the (CH₂)_(2n+1)—X—R6 group and PG is a protective group;    -   f) deprotecting the compound (Xl) in conditions known to those        skilled in the art.

According to a second embodiment, compounds represented by formula (I)according to the invention in which (i) G3 is an oxygen atom, (ii) G2represents a N—R4 group, (iii) R and R4 represent linear or branchedalkyl groups, saturated or 15 not, optionally substituted, containingfrom 1 to 5 carbon atoms, (iv) R1 and R3 are hydrogen atoms and (v) R2represents a CO—R5 group or a CO—(CH2)_(2n+1)—X—R6 group are obtained inthe following manner (diagram 4):

-   -   a) reacting a compound represented by formula (VIII) with a        compound PG′-E in which PG′ is a protective group and E is a        reactive group selected for example in the group consisting of        OH or a halogen, to give a compound represented by general        formula (XII) in which R represents a linear or branched alkyl        group, saturated or not, optionally substituted, containing from        1 to 5 carbon atoms and PG is another protective group such as        defined hereinabove. Advantageously, the reaction can be carried        out by adapting the protocols described by (Marx, Piantadosi et        al. 1988) and (Gaffney and Reese 1997) in which PG′-E can        represent triphenylmethyl chloride or 9-phenylxanthene-9-ol or        else 9-chloro-9-phenylxanthene;    -   b) reacting a compound represented by formula (XII) such as        defined hereinabove with a compound corresponding to the formula        LG-E in which E represents a halogen and LG is a reactive group        selected for example in the group consisting of mesyl, tosyl,        etc., to give a compound represented by general formula (XIII)        in which R represents a linear or branched alkyl group,        saturated or not, optionally substituted, containing from 1 to 5        carbon atoms and PG and PG′ are protective groups, by adapting        the method described by (Kitchin, Bethell et al. 1994);    -   c) reacting a compound represented by formula (XIII) such as        defined hereinabove with a compound corresponding to the formula        R4-NH₂ in which R4 represents a linear or branched alkyl group,        saturated or not, optionally substituted, containing from 1 to 5        carbon atoms and NH₂ represents the amine function, according to        the method described by (Ramalingan, Raju et al. 1995), to        obtain a compound represented by formula (XIV) in which R and R4        are independently such as defined hereinabove;    -   d) reacting a compound represented by formula (XIV) with a        compound corresponding to the formula A°-CO-A2 in which A2 is a        reactive group selected for example in the group consisting of        OH and Cl, and A° is the R5 group or the (CH₂)_(2n+1)—X—R6        group, possibly in the presence of coupling agents or activators        known to those skilled in the art to give a compound represented        by formula (XV) in which R and R4 independently represent linear        or branched alkyl groups, saturated or not, optionally        substituted, containing from 1 to 5 carbon atoms, R2 represents        a CO—R5 group or a CO—(CH₂)_(2n+1)—X—R6 group, PG and PG′ are        protective groups;    -   e) deprotecting a compound represented by formula (XV) in        conventional conditions known to those skilled in the art to        obtain a compound represented by general formula (I) according        to the invention in which (i) R and R4 independently represent        linear or branched alkyl groups, saturated or not, optionally        substituted, containing from 1 to 5 carbon atoms, (ii) R1 and R3        are hydrogen atoms and (iii) R2 represents a CO—R5 group or a        CO—(CH₂)_(2n+1)—X—R6 group.

Compounds represented by formula (I) according to the invention in which(i) G2 and G3 are sulfur atoms or a NH group, (ii) R is a hydrogen atomand (iii) R1, R2 and R3 are hydrogen atoms or represent a CO—R5 orCO—(CH₂)_(2n+1)—X—R6 group can be obtained by different methods.

According to a first embodiment, compounds represented by formula (I)according to the invention in which (i) G2 and G3 are sulfur atoms or aNH group, (ii) R is a hydrogen atom and (iii) R1, R2 and R3 are hydrogenatoms or represent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group, R1, R2 and/orR3 having the same meaning when they are carried on a same heteroatom(sulfur or nitrogen), can be obtained in the following manner (diagram5A):

-   -   a) reacting a compound represented by formula (IIa-c) with a        compound corresponding to the formula LG-E in which E represents        a halogen and LG is a reactive group selected for example in the        group consisting of mesyl, tosyl, etc., to give a compound        represented by general formula (XVIa-c);    -   b) reacting a compound represented by formula (XVIa-c) with a        compound corresponding to the formula Ac-S-B⁺ in which Ac        represents a short acyl group, preferably the acetyl group, and        B is a counter-ion selected for example in the group consisting        of sodium and potassium, preferably potassium to give the        compound represented by general formula (XVIIa-c).        Advantageously, said reaction can be carried out by adapting the        protocol described by (Gronowitz, Herslbf et al. 1978);    -   c) deprotecting a compound represented by formula (XVIla-c), in        conventional conditions known to those skilled in the art, and        for example in basic medium, to give a compound represented by        general formula (I) in which (i) G2 and G3 represent a sulfur        atom or a NH group and (ii) R1, R2 and R3, which are the same or        different, represent a hydrogen atom or a CO—R5 or        CO—(CH₂)_(2n+1)—X—R6 group;    -   d) reacting a compound represented by general formula (I) in        which (i) G2 and G3 represent a sulfur atom or a NH group        and (ii) R1, R2 and R3, which are the same or different,        represent a hydrogen atom or a CO—R5 or CO—(CH₂)_(2n+1)—X—R6        group, with a compound corresponding to the formula A°-CO-A2 in        which A2 is a reactive group selected for example in the group        consisting of OH and Cl, and A° is the R5 group or the        (CH₂)_(2n+1)—X—R6 group, possibly in the presence of coupling        agents or activators known to those skilled in the art.

According to similar synthetic method, compounds having formula (I)according to the invention in which (i) G2 and G3 are sulfur atoms or aNH group, (ii) R is a hydrogen atom and (iii) R1, R2 and R3 are hydrogenatoms or represent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group, R1, R2 and/orR3 having the same meaning when they are carried on a same heteroatom(sulfur or nitrogen), can be prepared in the following manner (diagram5B):

-   -   a) reacting a compound represented by formula (IIa-c) with a        compound corresponding to the formula (LG)2 in which LG is a        reactive group selected for example in the group consisting of        iodine, bromine, etc., possibly in the presence of activators        known to those skilled in the art to give a compound represented        by general formula (XVId-f);    -   b) reacting a compound represented by formula (XVld-f) with a        compound corresponding to the formula HS-B⁺ in which B is a        counter-ion selected for example in the group consisting of        sodium or potassium, preferably sodium to give a compound        represented by general formula (I) in which (i) G2 and G3        represent a sulfur atom or a NH group and (ii) R1, R2 and R3,        which are the same or different, represent a hydrogen atom or a        CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group;    -   c) reacting a compound represented by general formula (I) in        which (i) G2 and G3 represent a sulfur atom or a NH group        and (ii) R1, R2 and R3, which are the same or different,        represent a hydrogen atom or a CO—R5 or CO—(CH₂)_(2n+1)—X—R6        group, with a compound corresponding to the formula A°-CO-A2 in        which A2 is a reactive group selected for example in the group        consisting of OH and Cl, and A° is the R5 group or the        (CH₂)_(2n+1)—X—R6 group, possibly in the presence of coupling        agents or activators known to those skilled in the art.

Said reaction enables the synthesis of compounds represented by generalformula (I) in which the groups carried on a same heteroatom (nitrogenor sulfur) respectively (R2 and R3), (R1 and R3) and (R1 and R2) havethe same meaning.

The above steps can be carried out in an advantageous manner accordingto the protocols described by (Adams, Doyle et al. 1960) and (Gronowitz,Herslbf et al. 1978).

According to another method of the invention (diagram 6), compoundsrepresented by formula (I) according to the invention in which (i) G2and G3 are sulfur atoms or a NH group, (ii) R is a hydrogen atom and(iii) R1, R2 and R3 are hydrogen atoms or represent a CO—R5 orCO—(CH₂)_(2n+1)—X—R6 group can be prepared from compounds represented byformula (IIIa-c) by a method comprising:

-   -   a) reacting a compound represented by formula (illa-c) with a        compound corresponding to the formula LG-E in which E represents        a halogen and LG is a reactive group selected for example in the        group consisting of mesyl, tosyl, etc., to give a compound        represented by general formula (XVIIIa-c) in which PG represents        a protective group;    -   b) reacting a compound represented by formula (XVIIIa-c) with a        compound corresponding to the formula Ac-S-B⁺ in which Ac        represents a short acyl group, preferably the acetyl group, and        B is a counter-ion selected for example in the group consisting        of sodium and potassium, preferably potassium to give a compound        represented by general formula (XIXa-c). Advantageously, said        reaction can be carried out by adapting the protocol described        by (Gronowitz, Herslbf et al. 1978);    -   c) deprotecting the sulfur atom of a compound (XIXa-c) in        conditions known to those skilled in the art, to give a compound        represented by general formula (XXa-c);    -   d) reacting a compound represented by general formula (XXa-c)        with a compound corresponding to the formula A°-CO-A2 in which        A2 is a reactive group selected for example in the group        consisting of OH and Cl, and A° is the R5 group or the        (CH₂)_(2n+1)—X—R6 group, possibly in the presence of coupling        agents or activators known to those skilled in the art to give a        compound represented by general formula (XXla-c) in which R2 and        R3 represent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group;    -   e) deprotecting a compound represented by formula (XXIa-c) in        conventional conditions known to those skilled in the art, to        give a compound represented by formula (I) according to the        invention in which (i) G2 and G3 are sulfur atoms or a NH        group, (ii) R and R1 are hydrogen atoms and (iii) R2 and R3        represent a hydrogen atom, a CO—R5 or CO—(CH₂)_(2n+1)—X—R6        group.    -   f) reacting a compound represented by formula (I) according to        the invention in which (i) G2 and G3 are sulfur atoms or a NH        group, (ii) R and R1 are hydrogen atoms and (iii) R2 and R3        represent a hydrogen atom, a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group        with a compound corresponding to the formula A°-CO-A2 in which        A2 is a reactive group selected for example in the group        consisting of OH and Cl, and A° is the R5 group or the        (CH₂)_(2n+1)—X—R6 group, possibly in the presence of coupling        agents or activators known to those skilled in the art.

Said reaction enables the synthesis of compounds represented by generalformula (I) in which the groups carried on a same heteroatom (nitrogenor sulfur) respectively (R2 and R3), (R1 and R3) and (R1 and R2) havethe same meaning.

In an advantageous manner, the above steps are carried out according tothe protocols described by (Adams, Doyle et al. 1960), (Gronowitz,Herslbf et al. 1978), (Bhatia and Hajdu 1987) and (Murata, Ikoma et al.1991).

Compounds represented by general formula (I) in which (i) G2 and G3represent sulfur atoms or a N—R4 group, (ii) R and R4 independentlyrepresent a linear or branched alkyl group, saturated or not, optionallysubstituted, containing from 1 to 5 carbon atoms, (iii) R1, R2 and R3,which are the same or different, represent a CO—R5 group or aCO—(CH₂)_(2n+1)—X—R6 group, are obtained by reacting a compoundrepresented by general formula (I) in which (i) G2 or G3 represent asulfur atom or a N—R4 group, (ii) R and R4 independently representgroups such as defined hereinabove, (iii) R1 is a hydrogen atom and (iv)R2 and R3, which are the same or different, represent a CO—R5 group or aCO—(CH₂)_(2n+1)—X—R6 group with a compound corresponding to the formulaA°-CO-A2 in which A2 is a reactive group selected for example in thegroup consisting of OH and Cl, and A° is the R5 group or the(CH₂)_(2n+1)—X—R6 group, possibly in the presence of coupling agents oractivators known to those skilled in the art.

Compounds represented by general formula (I) in which (i) the group G2and G3 represent sulfur atoms or a N—R4 group, (ii) R and R4independently represent groups such as defined hereinabove, (iii) R1 isa hydrogen atom and (iv) R2 and R3, which are the same or different,represent a CO—R5 group or a CO—(CH₂)_(2n+1)—X—R6 group, can be obtainedby the following methods:

In a first embodiment, compounds represented by formula (I) according tothe invention in which (i) the group G2 is a sulfur atom, (ii) G3represents a N—R4 group, (iii) R and R4 independently representdifferent linear or branched alkyl group, saturated or not, optionallysubstituted, containing from 1 to 5 carbon atoms, (iv) R1 is a hydrogenatom and (v) R2 and R3, which are the same or different, represent aCO—R5 group or a CO—(CH₂)_(2n+1)—X—R6 group are obtained in thefollowing manner (diagram 7):

-   -   a) reacting a compound represented by formula (XI) with a        compound corresponding to the formula LG-E in which E represents        a halogen and LG is a reactive group selected for example in the        group consisting of mesyl, tosyl, etc., to give a compound        represented by general formula (XXII) in which PG represents a        protective group;    -   b) reacting a compound represented by formula (XXII) with a        compound corresponding to the formula Ac-S-B⁺ in which Ac        represents a short acyl group, preferably the acetyl group, and        B is a counter-ion selected for example in the group consisting        of sodium and potassium, preferably potassium to give the        compound represented by general formula (XXIII). Advantageously,        said reaction is carried out by adapting the protocol described        by (Gronowitz, Herslöf et al. 1978);    -   c) deprotecting the sulfur atom of a compound represented by        formula (XXIII) in conventional conditions known to those        skilled in the art to give a compound represented by general        formula (XXIV);    -   d) reacting a compound represented by general formula (XXIV)        with a compound corresponding to the formula A°-CO-A2 in which        A2 is a reactive group selected for example in the group        consisting of OH and Cl, and A° is the R5 group or the        (CH₂)_(2n+1)—X—R6 group, possibly in the presence of coupling        agents or activators known to those skilled in the art to give a        compound represented by general formula (XXV) in which R2 and        R3, which are the same or different, represent a CO—R5 or        CO—(CH₂)_(2n+1)—X—R6 group;    -   e) deprotectig the compound of formula (XXV) in conditions known        to those skilled in the art.

According to another method (diagram 8), compounds represented byformula (I) according to the invention in which (i) G2 represents a N—R4group, (ii) G3 is a sulfur atom, (iii) R and R4 independently representdifferent linear or branched alkyl groups, saturated or not, optionallysubstituted, containing from 1 to 5 carbon atoms, (iv) R1 is a hydrogenatom and (v) R2 and R3, which are the same or different, represent aCO—R5 group or a CO—(CH₂)_(2n+1)—X—R6 group are obtained in thefollowing manner:

-   -   a) reacting the compound represented by formula (IX) with a        compound corresponding to the formula Ac-S-B⁺ in which Ac        represents a short acyl group, preferably the acetyl group, and        B is a counter-ion selected for example in the group consisting        of sodium and potassium, preferably potassium to give the        compound represented by general formula (XXVI). Advantageously,        said reaction can be carried out by adapting the protocol        described by (Gronowitz, Herslöf et al. 1978);    -   b) reacting a compound represented by formula (XXVI) with a        compound corresponding to the formula LG-E in which E represents        a halogen and LG is a reactive group selected for example in the        group consisting of mesyl, tosyl, etc., to give a compound        represented by general formula (XXVII) in which PG represents a        protective group;    -   c) reacting the compound (XXVII) with a compound represented by        formula R4-NH₂ in which R4 represents a linear or branched alkyl        group, saturated or not, optionally substituted, containing from        1 to 5 carbon atoms and NH₂ represents the amine function,        according to the method described by (Ramalingan, Raju et al.        1995), to give a compound represented by formula (XXVIII) in        which R and R4 independently represent different linear or        branched alkyl groups, saturated or not, optionally substituted,        containing from 1 to 5 carbon atoms;    -   d) reacting a compound represented by general formula (XXVIII)        with a compound corresponding to the formula A°-CO-A2 in which        A2 is a reactive group selected for example in the group        consisting of OH and Cl, and A° is the R5 group or the        (CH₂)_(2n+1)—X—R6 group, possibly in the presence of coupling        agents or activators known to those skilled in the art to give a        compound represented by general formula (XXIX);    -   e) deprotecting the sulfur atom of a compound represented by        formula (XXIX) in conventional conditions known to those skilled        in the art to give a compound represented by general formula        (XXX);    -   f) reacting a compound represented by general formula (XXX) with        a compound corresponding to the formula A°-CO-A2 in which A2 is        a reactive group selected for example in the group consisting of        OH and Cl, and A° is the R5 group or the (CH₂)_(2n+1)—X—R6        group, possibly in the presence of coupling agents or activators        known to those skilled in the art to give a compound represented        by general formula (XXXI) in which R2 and R3, which are the same        or different, represent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group;    -   g) deprotecting a compound represented by formula (XXXI) in        conventional conditions known to those skilled in the art.

Compounds represented by formula (I) according to the invention in which(i) G2 is a sulfur atom, (ii) G3 is an oxygen atom, (iii) R is ahydrogen atom, (iv) R1 and R2 represent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6group and (v) R3 is a hydrogen atom or represents a CO—R5 orCO—(CH₂)_(2n+1)—X—R6 group, can be prepared from compounds havingformula (V) according to the following method (diagram 9A):

-   -   a) reacting the compound (V) with a compound corresponding to        the formula LG-E in which E represents a halogen and LG is a        reactive group selected for example in the group consisting of        mesyl, tosyl, etc., to give a compound represented by general        formula (XXXII) in which PG represents a protective group;    -   b) reacting a compound represented by formula (XXXII) with a        compound corresponding to the formula Ac-S-B⁺ in which Ac        represents a short acyl group, preferably the acetyl group, and        B is a counter-ion selected for example in the group consisting        of sodium and potassium, preferably potassium to give the        compound represented by general formula (XXXIII).        Advantageously, said reaction can be carried out by adapting the        protocol described by (Gronowitz, Herslof et al. 1978);    -   c) deprotecting the sulfur atom of a compound (XXXIII), in        conventional conditions known to those skilled in the art, to        give a compound represented by general formula (XXXIV);    -   d) reacting a compound represented by general formula (XXXIV)        with a compound corresponding to the formula A°-CO-A2 in which        A2 is a reactive group selected for example in the group        consisting of OH and Cl, and A° is the R5 group or the        (CH₂)_(2n+1)—X—R6 group, possibly in the presence of coupling        agents or activators known to those skilled in the art to give a        compound represented by general formula (XXXV) in which R1 and        R2, which are the same or different, represent a CO—R5 or        CO—(CH₂)_(2n+1)—X—R6 group;    -   e) deprotecting a compound (XXXV) in conventional conditions        known to those skilled in the art to give a compound represented        by general formula (I) in which G2 is a sulfur atom, G3 is an        oxygen atom, R and R3 are hydrogen atoms and R1 and R2, which        are the same or different, represent a CO—R5 or        CO—(CH₂)_(2n+1)—X—R6 group;    -   f) reacting a compound represented by general formula (I) in        which (i) G2 is a sulfur atom, (ii) G3 is an oxygen atom, (iii)        R and R3 are hydrogen atoms and (iv) R1 and R2, which are the        same or different, represent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6        group with a compound corresponding to the formula A°-CO-A2 in        which A2 is a reactive group selected for example in the group        consisting of OH and Cl, and A° is the R5 group or the        (CH₂)_(2n+1)—X—R6 group, possibly in the presence of coupling        agents or activators known to those skilled in the art.

According to a similar method of synthesis, compounds represented byformula (I) according to the invention in which (i) G2 is a sulfur atom,(ii) G3 is an oxygen atom, (iii) R is a hydrogen atom, (iv) R1 and R2represent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group and (v) R3 is a hydrogenatom or represents a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group, can beprepared from compounds of formula (V) by the following method (diagram9B):

-   -   a) reacting the compound (V) with a compound corresponding to        the formula (LG)₂ in which LG is a reactive group selected for        example in the group consisting of iodine, bromine, etc., to        give a compound represented by general formula (XXXIIa) in which        PG represents a protective group;    -   b) reacting a compound represented by formula (XXXIIa) with a        compound corresponding to the formula HS-B+in which B is a        counter-ion selected for example in the group consisting of        sodium and potassium, preferably sodium to give a compound        represented by general formula (XXXIV);    -   c) reacting a compound represented by general formula (XXXIV)        with a compound corresponding to the formula A°-CO-A2 in which        A2 is a reactive group selected for example in the group        consisting of OH and Cl, and A° is the R5 group or the        (CH₂)_(2n+1)—X—R6 group, possibly in the presence of coupling        agents or activators known to those skilled in the art to give a        compound represented by general formula (XXXV) in which R1 and        R2, which are the same or different, represent a CO—R5 or        CO—(CH₂)_(2n+1)—X—R6 group;    -   d) deprotecting the compound (XXXV) in conventional conditions        known to those skilled in the art to give a compound represented        by general formula (I) in which G2 is a sulfur atom, G3 is an        oxygen atom, R and R3 are hydrogen atoms and R1 and R2, which        are the same or different, represent a CO—R5 or        CO—(CH₂)_(2n+1)—X—R6 group;    -   e) reacting a compound represented by general formula (I) in        which (i) G2 is a sulfur atom, (ii) G3 is an oxygen atom, (iii)        R and R3 are hydrogen atoms and (iv) R1 and R2, which are the        same or different, represent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6        group with a compound corresponding to the formula A°-CO-A2 in        which A2 is a reactive group selected for example in the group        consisting of OH and Cl, and A° is the R5 group or the        (CH₂)_(2n+1)—X—R6 group, possibly in the presence of coupling        agents or activators known to those skilled in the art.

Compounds represented by formula (I) according to the invention in which(i) G2 is a sulfur atom, (ii) G3 is an oxygen atom, (iii) R is ahydrogen atom, (iv) R1 and R3 represent a hydrogen atom or a CO—R5 orCO—(CH₂)_(2n+1)—X—R6 group, which are the same or different, and (v) R2represents a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group, can be prepared fromcompounds having formula (IIIa) by the following method (diagram 10):

-   -   a) reacting a compound represented by formula (IIIa) with a        compound PG′-E in which PG′ is a protective group and E is a        reactive group selected for example in the group consisting of        OH or a halogen, to give a compound represented by general        formula (XXXVI) in which PG is another protective group such as        defined earlier. In an advantageous manner, the reaction can be        carried out by adapting the protocols described by (Marx,        Piantadosi et al. 1988) and (Gaffney and Reese 1997) in which        PG-E can represent triphenylmethyl chloride or        9-phenylxanthene-9-ol or else 9-chloro-9-phenylxanthene;    -   b) reacting the compound (XXXVI) with a compound corresponding        to the formula LG-E in which E represents a halogen and LG is a        reactive group selected for example in the group consisting of        mesyl, tosyl, etc., to give a compound represented by general        formula (XXXVII) in which PG and PG′ represent judiciously        selected protective groups such as defined hereinabove;    -   c) reacting a compound represented by formula (XXXVII) with a        compound corresponding to the formula Ac-S-B+in which Ac        represents a short acyl group, preferably the acetyl group, and        B is a counter-ion selected for example in the group consisting        of sodium and potassium, preferably potassium to give the        compound represented by general formula (XXXVIII).        Advantageously, said reaction can be carried out by adapting the        protocol described by (Gronowitz, Herslbf et al. 1978);    -   d) deprotecting the sulfur atom of a compound (XXXVIII), in        conventional conditions known to those skilled in the art, to        give a compound represented by general formula (XXXIX);    -   e) reacting a compound represented by general formula (XXXIX)        with a compound corresponding to the formula A°-CO-A2 in which        A2 is a reactive group selected for example in the group        consisting of OH and Cl, and A° is the R5 group or the        (CH₂)_(2n+1)—X—R6 group, possibly in the presence of coupling        agents or activators known to those skilled in the art to give a        compound represented by general formula (XL) in which R2        represents a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group;    -   f) deprotecting a compound (XL) in conventional conditions known        to those skilled in the art to give a compound represented by        general formula (I) in which G2 is a sulfur atom, G3 is an        oxygen atom, R, R1 and R3 are hydrogen atoms and R2 represents a        CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group (compound XLI);    -   g) reacting a compound represented by formula (XLI) with a        compound (PG)₂O in which PG is a protective group to give a        compound represented by general formula (XLII). Advantageously,        the reaction can be carried out by adapting the protocols        described by (Nazih, Cordier et al. 2000) and (Kotsovolou, Chiou        et al. 2001) in which (PG)₂O represents di-tert-butyl        dicarbonate;    -   h) reacting a compound represented by general formula (XLII)        with a compound corresponding to the formula A°-CO-A2 in which        A2 is a reactive group selected for example in the group        consisting of OH and Cl, and A° is the R5 group or the        (CH₂)_(2n+1)—X—R6 group, possibly in the presence of coupling        agents or activators known to those skilled in the art to give a        compound represented by formula (XLIII);    -   i) deprotecting a compound (XLIII) in conventional conditions        known to those skilled in the art to give a compound represented        by general formula (I) in which G2 is a sulfur atom, G3 is an        oxygen atom, R and R1 are hydrogen atoms and R2 and R3, which        are the same or different, represent a CO—R5 or        CO—(CH₂)_(2n+1)—X—R6 group;    -   j) reacting a compound represented by general formula (I) in        which G2 is a sulfur atom, G3 is an oxygen atom, R and R1 are        hydrogen atoms and R2 and R3, which are the same or different,        represent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group with a compound        corresponding to the formula A°-CO-A2 in which A2 is a reactive        group selected for example in the group consisting of OH and Cl,        and A° is the R5 group or the (CH₂)_(2n+1)—X—R6 group, possibly        in the presence of coupling agents or activators known to those        skilled in the art.

Compounds represented by formula (I) according to the invention in which(i) G2 is an oxygen atom, (ii) G3 is a sulfur atom, (iii) R is ahydrogen atom, (iv) R1 and R3 are hydrogen atoms or represent a CO—R5 orCO—(CH₂)_(2n+1)—X—R6 group and (v) R2 represents a hydrogen atom or aCO—R5 or CO—(CH₂)_(2n+1)—X—R6 group, can be prepared from compoundshaving formula (IIa) according to the following method (diagram 11):

-   -   a) reacting a compound represented by formula (IIa) such as        defined hereinabove, with a compound corresponding to the        formula LG-E (in stoichiometric amounts) in which E represents a        halogen and LG is a reactive group selected for example in the        group consisting of mesyl, tosyl, etc., to give a compound        represented by general formula (XLIV);    -   b) reacting a compound represented by formula (XLIV) with a        compound corresponding to the formula Ac-S-B⁺ in which Ac        represents a short acyl group, preferably the acetyl group, and        B is a counter-ion selected for example in the group consisting        of sodium and potassium, preferably potassium to give the        compound represented by general formula (XLV). Advantageously,        said reaction can be carried out by adapting the protocol        described by (Gronowitz, Herslöf et al. 1978);    -   c) reacting a compound represented by formula (XLV) with a        compound PG-E in which PG is a protective group and E is a        reactive group selected for example in the group consisting of        OH and a halogen, to give a compound represented by general        formula (XLVI). Advantageously, the reaction can be carried out        by adapting the protocols described by (Marx, Piantadosi et        al. 1988) and (Gaffney and Reese 1997), in which PG-E can        represent triphenylmethyl chloride or 9-phenylxanthene-9-ol or        else 9-chloro-9-phenylxanthene;    -   d) deprotecting the sulfur atom of a compound (XLVI), in        conditions known to those skilled in the art, to give a compound        represented by general formula (XLVII);    -   e) reacting a compound represented by general formula (XLVII)        with a compound corresponding to the formula A°-CO-A2 in which        A2 is a reactive group selected for example in the group        consisting of OH and Cl, and A° is the R5 group or the        (CH₂)_(2n+1)—X—R6 group, possibly in the presence of coupling        agents or activators known to those skilled in the art to give a        compound represented by general formula (XLVIII) in which R1 and        R3, which are the same or different, represent a CO—R5 or        CO—(CH₂)_(2n+1)—X—R6 group;    -   f) deprotecting a compound represented by formula (XLVIII), in        conventional conditions known to those skilled in the art, to        give a compound represented by general formula (I) in which G2        is an oxygen atom, G3 is a sulfur atom, R and R2 are hydrogen        atoms and R1 and R3, which are the same or different, represent        a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group;    -   g) reacting a compound represented by general formula (I) in        which G2 is an oxygen atom, G3 is a sulfur atom, R and R2 are        hydrogen atoms and R1 and R3, which are the same or different,        represent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group with a compound        corresponding to the formula A°-CO-A2 in which A2 is a reactive        group selected for example in the group consisting of OH and Cl,        and A° is the R5 group or the (CH₂)_(2n+1)—X—R6 group, possibly        in the presence of coupling agents or activators known to those        skilled in the art.

Compounds represented by formula (I) according to the invention in which(i) G2 is an oxygen atom, (ii) G3 is a sulfur atom, (iii) R is ahydrogen atom, (iv) R1 and R3 are hydrogen atoms or represent a CO—R5 orCO—(CH₂)_(2n+1)—X—R6 group, which are the same or different, and (v) R3represents a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group, can be rpepared fromcompounds having formula (IIIa) according to the following method(diagram 12):

-   -   a) reacting a compound represented by formula (IIIa) such as        defined hereinabove, with a compound corresponding to the        formula LG-E (in stoichiometric amounts) in which E represents a        halogen and LG is a reactive group selected for example in the        group consisting of mesyl, tosyl, etc., to give a compound        represented by general formula (XLIX);    -   b) reacting a compound represented by formula (XLIX) with a        compound corresponding to the formula Ac-S-B⁺ in which Ac        represents a short acyl group, preferably the acetyl group, and        B is a counter-ion selected for example in the group consisting        of sodium and potassium, preferably potassium to give the        compound represented by general formula (L). Advantageously,        said reactino can be carried out by adapting the protocol        described by (Gronowitz, Herslof et al. 1978);    -   c) reacting a compound represented by formula (L) with a        compound PG′-E in which PG′ is a protective group and E is a        reactive group selected for example in the group consisting of        OH and a halogen, to give a compound represented by general        formula (LI). Advantageously, the reaction can be carried out by        adapting the protocols described by (Marx, Piantadosi et        al. 1988) and (Gaffney and Reese 1997) in which PG′-E can        represent triphenylmethyl chloride or 9-phenylxanthene-9-ol or        else 9-chloro-9-phenylxanthene;    -   d) deprotecting the sulfur atom of a compound (LI), in        conditions known to those skilled in the art, to give a compound        represented by general formula (LII);    -   e) reacting a compound represented by general formula (LII) with        a compound corresponding to the formula A°-CO-A2 in which A2 is        a reactive group selected for example in the group consisting of        OH and Cl, and A° is the R5 group or the (CH₂)_(2n+1)—X—R6        group, possibly in the presence of coupling agents or activators        known to those skilled in the art to give a compound represented        by general formula (LIII) in which R3 represents a CO—R5 or        CO—(CH₂)_(2n+1)—X—R6 group;    -   f) deprotecting a compound represented by formula (LIII), in        conventional conditions known to those skilled in the art, to        give a compound represented by general formula (I) in which G2        is an oxygen atom, G3 is a sulfur atom, R and R2 are hydrogen        atoms and R3 represents a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group        (compound LIV);    -   g) reacting a compound represented by formula (LIV) with a        compound (PG)₂O in which PG is a protective group to give a        compound represented by general formula (LV). Advantageously,        the reaction can be carried out by adapting the protocols        described by (Nazih, Cordier et al. 2000) and (Kotsovolou, Chiou        et al. 2001) in which (PG)₂O represents di-tert-butyl        dicarbonate;    -   h) reacting a compound represented by general formula (LV) with        a compound corresponding to the formula A°-CO-A2 in which A2 is        a reactive group selected for example in the group consisting of        OH and Cl, and A° is the R5 group or the (CH₂)_(2n+1)—X—R6        group, possibly in the presence of coupling agents or activators        known to those skilled in the art to give a compound represented        by formula (LVI);    -   i) deprotecting a compound (LVI) in conventional conditions        known to those skilled in the art to give a compound represented        by general formula (I) in which G3 is a sulfur atom, G2 is an        oxygen atom, R and R1 are hydrogen atoms and R2 and R3, which        are the same or different, represent a CO—R5 or        CO—(CH₂)_(2n+1)—X—R6 group;    -   j) reacting a compound represented by general formula (I) in        which G3 is a sulfur atom, G2 is an oxygen atom, R and R1 are        hydrogen atoms and R2 and R3, which are the same or different,        represent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group with a compound        corresponding to the formula A°-CO-A2 in which A2 is a reactive        group selected for example in the group consisting of OH and Cl,        and A° is the R5 group or the (CH₂)_(2n+1)—X—R6 group, possibly        in the presence of coupling agents or activators known to those        skilled in the art.

Compounds represented by formula (I) according to the invention in which(i) G2 is an oxygen atom, (ii) G3 is a sulfur atom, (iii) R is ahydrogen atom, (iv) R2 and R3, which are the same, are hydrogen atoms orrepresent a CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group and (v) R1 represents aCO—R5 or CO—(CH₂)_(2n+1)—X—R6 group, can be prepared from compoundshaving formula (IIIa) according to the following method (diagram 13):

-   -   a) reacting a compound represented by formula (IIIa) such as        defined hereinabove, with a compound corresponding to the        formula (LG)₂ (in stoichiometric amounts) in which LG is a        reactive group selected for example in the group consisting of        iodine, bromine, etc., to give a compound represented by general        formula (XLIXa);    -   b) reacting a compound represented by formula (XLIXa) with a        compound corresponding to the formula Ac-S-B⁺ in which Ac        represents a short acyl group, preferably the acetyl group, and        B is a counter-ion selected for example in the group consisting        of sodium and potassium, preferably potassium to give the        compound represented by general formula (L);    -   c) deprotecting the sulfur atom of a compound (L), in conditions        known to those skilled in the art, to give a compound        represented by general formula (LVII);    -   d) reacting a compound represented by general formula (LVII)        with a compound corresponding to the formula A°-CO-A2 in which        A2 is a reactive group selected for example in the group        consisting of OH and Cl, and A° is the R5 group or the        (CH₂)_(2n+1)—X—R6 group, possibly in the presence of coupling        agents or activators known to those skilled in the art to give a        compound represented by general formula (LVI) in which R2 and R3        represent a same CO—R5 or CO—(CH₂)_(2n+1)—X—R6 group;    -   e) deprotecting a compound represented by formula (LVI), in        conventional conditions known to those skilled in the art, to        give a compound represented by general formula (I) in which G2        is an oxygen atom, G3 is a sulfur atom, R and R2 are hydrogen        atoms and R2 and R3 represent a same CO—R5 or        CO—(CH₂)_(2n+1)—X—R6 group;    -   f) reacting a compound represented by general formula (I) in        which G2 is an oxygen atom, G3 is a sulfur atom, R and R2 are        hydrogen atoms and R2 and R3 represent a same CO—R5 or        CO—(CH₂)_(2n+1)—X-R6 group with a compound corresponding to the        formula A°-CO-A2 in which A2 is a reactive group selected for        example in the group consisting of OH and Cl, and A° is the R5        group or the (CH₂)_(2n+1)—X—R6 group, possibly in the presence        of coupling agents or activators known to those skilled in the        art.

The feasibility, realization and other advantages of the invention arefurther detailed in the following examples, which are given for purposesof illustration and not by way of limitation.

Legends of Figures:

FIG. 1: Structure of particular inventive compounds the preparation ofwhich is described in examples 2, 4, 5, 6, 8, 10 to 14, 16, 18, 19, 21and 23 and respectively noted on the figure as 1A.2, 1A.4, 1A.5, 1A.6,1A.8, 1A.10, 1A.11, 1A.12, 1A.13, 1A.14, 1A.16, 1A.18, 1A.19, 1A.21 and1A.23.

FIG. 2: Evaluation of the antioxidant properties of the inventivecompounds on LDL oxidation by copper (Cu).

-   -   FIG. 2 a: conjugated diene formation over time or lag phase.    -   FIG. 2 b: rate of diene formation.    -   FIG. 2 c: maximum amount of conjugated dienes formed.

FIG. 3: Evaluation of the PPAR□ agonist properties of the inventivecompounds with the Gal4/PPAR□ transactivation system.

EXAMPLES

For easier comprehension of the text, the inventive compounds used inthe examples concerning the measurement and evaluation of activity areabbreviated as follows: “Ex 2”, for instance, indicates the compound ofthe invention which preparation is described in example 2.

Thin-layer chromatography (TLC) was carried out on plates coated withMERCK silica gel 60F₂₅₄ 0.2 mm thick. Retention factor is abbreviatedRf.

Column chromatography was carried out on silica gel 60 with a particlesize of 40-63 μm (Merck reference 9385-5000).

Melting points (MP) were determined on a Buchi B 540 apparatus by thecapillary method.

Infrared (IR) spectra were recorded on a Bruker Fourier transformationspectrometer (Vector 22).

Nuclear magnetic resonance (NMR) spectra were recorded on a Bruker AC300 spectrometer (300 MHz). Each signal was identified by its chemicalshift, intensity, multiplicity (noted s for singlet, sl for broadsinglet, d for doublet, dd for split doublet, t for triplet, td forsplit triplet, quint for quintuplet and m for multiplet) and itscoupling constant (J).

Mass spectra (MS) were determined on a Perkin Elmer Sciex API 1 (ESI-MSfor ElectroSpray Ionization Mass Spectrometry) or on an AppliedBiosystems Voyager DE-STR of the MALDI-TOF type (Matrix-Assisted LaserDesorption/Ionization—Time Of Flight).

Example 1 Preparation of tetradecylthioacetic Acid

Potassium hydroxide (34.30 g, 0.611 mol), mercaptoacetic acid (20.9 ml,0.294 mol) and 1-bromotetradecane (50 ml, 0.184 mol) were added in thatorder to methanol (400 ml). The mixture was stirred overnight at roomtemperature. A concentrated hydrochloric acid solution (60 ml) dissolvedin water (800 ml) was then added. The tetradecylthioacetic acidprecipitated. The mixture was stirred overnight at room temperature. Theprecipitate was then filtered, washed five times with water and dried ina dessicator. The product was recrystallized in methanol.

Yield: 94%

Rf (dichloromethane/methanol 9:1): 0.60

MP: 67-68° C.

IR: νCO acid 1726 and 1684 cm⁻¹

NMR (¹H, CDCl₃): 0.84-0.95 (t, 3H, —CH₃, J=6.5 Hz); 1.20-1.45(multiplet, 22H, —CH₂—); 1.55-1.69 (quint, 2H, —CH₂—CH₂—S—, J=7 Hz);2.63-2.72 (t, 2H, CH₂—CH₂—S—, J=7 Hz); 3.27 (s, 2H, S—CH₂—COOH)

MS (ESI-MS): M-1=287.

Example 2 Preparation of 3-(tetradecylthioacetylamino)propane-1.2-diol

Tetradecylthioacetic acid (example 1) (14.393 g, 50 mmol) and3-amino-propane-1,2-diol (5 g, 55 mmol) were placed in a flask andheated at 190° C. for 1 hour. The reaction mixture was cooled to roomtemperature, taken up in chloroform and washed once with water. Theorganic phase was dried on magnesium sulfuate, filtered and dried. Theresidue was stirred in ether and the product was isolated by filtration.

Yield: 22%

Rf (dichloromethane/methanol 9:1): 0.60

MP: 89-92° C.

IR: νNH and OH 3282 cm⁻¹; νCO amide 1640 cm⁻¹

NMR (¹H, CDCl₃): 0.89 (t, 3H, —CH₃, J=6.5 Hz); 1.26 (multiplet, 22H,—CH₂—); 1.57 (m, 2H, —CH₂—CH₂—S—); 2.54 (t, 2H, —CH₂—CH₂—S—, J=7.6 Hz);3.27 (s, 2H, S—CH₂—CONH—); 3.47 (m, 2H, —CONH—CH₂—CHOH—CH₂OH); 3.58 (m,1H, —CONH—CH₂—CHOH—CH₂OH); 3.81 (m, 2H, —CONH—CH₂—CHOH—CH₂OH); 7.33 (sl,1H, —CONH).

MS (MALDI-TOF): M+1=362 (M+H); M+23=385 (M+Na⁺); M+39=400 (M+K⁺)

Example 3 3-(Palmitoylamino)propane-1.2-diol

This compound was synthesized according to the method describedhereinabove (example 2) from 3-amino-propane-1,2-diol and palmitic acid.

Yield: 86%

Rf (dichloromethane/methanol 9:1): 0.50

IR: vNH and OH 3312 cm⁻¹; vCO amide 1633 cm⁻¹

MP: 104-108° C.

NMR (¹H, CDCl₃): 0.89 (t, 3H, —CH₃, J=6.5 Hz); 1.28 (multiplet, 24H,—CH₂—); 1.64 (m, 2H, —CH₂—CH₂-CO—); 2.24 (m, 2H, —CH₂—CH₂-CO—); 3.43 (m,2H, —CONH—CH₂—CHOH—CH₂OH); 3.55 (m, 2H, —CONH—CH₂—CHOH—CH₂OH); 3.78(m,1H, —CONH—CH₂—CHOH—CH₂OH); 5.82 (sl, 1H, —CONH—). MS (MALDI-TOF):M+1=330 (M+H)

Example 4 Preparation of1.2-(diPalmitoyloxy)-3-tetradecylthioacetylaminopropane

3-(tetradecylthioacetylamino)propane-1,2-diol (example 2) (1 g, 2.77mmol) was dissolved in dichloromethane (200 ml).Dicyclohexylcarbodiimide (1.426 g, 6.91 mmol), dimethylaminopyridine(0.845 g, 6.91 mmol) and palmitic acid (1.773 g, 6.91 mmol) were thenadded and the mixture was stirred at room temperature for 48 hours. Thedicyclohexylurea which precipitated was filtered and washed withdichloromethane. The filtrate was vacuum evaporated. The residue waspurified by chromatography on silica gel (eluent:dichloromethane/cyclohexane 6:4) (yield: 28%).

Rf (dichloromethane/cyclohexane 7:3): 0.28

MP: 73-75° C.

IR: νNH 3295 cm⁻¹; νCO ester 1730 cm⁻¹; vCO amide 1663 cm⁻¹ NMR (1H,CDC₃): 0.89 (t, 9H, —CH₃, J=6.5 Hz); 1.26 (multiplet, 70H, —CH₂—); 1.57(multiplet, 6H, —CH₂—CH₂—S— and OCOCH₂—CH₂); 2.33 (t, 4H, OCOCH₂—CH₂—,J=7.3 Hz); 2.51 (t, 2H, CH₂—CH₂—S—, J=7.3 Hz); 3.22 (s, 2H,S—CH₂—CONH—); 3.47 (m, 1H, —CONH—CHaHb-CH—CHcHd-); 3.62 (m, 1H,—CONH—CHaHb-CH—CHcHd); 4.12 (dd, 1H, —CHaHb-CH—CHcHd-, J=12.1 Hz andJ=5.7 Hz); 4.36 (dd, 1H, —CHaHb-CH—CHcHd-, J=12.1 Hz and J=4.4 Hz); 5.15(m, 1H, —CHaHb-CH—CHaHb); 7.20 (m, 1H, —NHCO—).

MS (MALDI-TOF): M+1=838 (M+H); M+23=860 (M+Na⁺); M+39=876 (M+K⁺)

Example 5 Preparation of1.2-(ditetradecylthioacetyloxy)-3-tetradecylthioacetylaminopropane

This compound was synthesized according to the method describedhereinabove (example 4) from3-(tetradecylthioacetylamino)propane-1,2-diol (example 2) andtetradecylthioacetic acid (example 1).

Yield: 41%

Rf (dichloromethane): 0.23

IR: νNH 3308 cm⁻¹; νCO ester 1722 and 1730 cm⁻¹; νCO amide 1672 cm⁻¹

MP: 65-67° C.

NMR (¹H, CDCl₃): 0.89 (t, 9H, —CH₃, J=6.4 Hz); 1.26 (multiplet, 66H,—CH₂—); 1.59 (multiplet, 6H, —CH₂—CH₂—S—); 2.53 (t, 2H,—CH₂—CH₂—S—CH₂—CONH—, J=7.3 Hz); 2.64 (t, 4H, CH₂—CH₂—S—CH₂—COO—, J=7.3Hz); 3.23 (s, 4H, S—CH₂—COO—); 3.24 (s, 2H, S—CH₂—CONH—); 3.52 (m, 1H,—CONH—CHaHb-CH—CHcHd-); 3.67 (m, 1H, —CONH—CHaHb-CH—CHcHd-); 4.22 (dd,1H, —CHaHb-CH—CHcHd-, J=12.2 Hz and J=5.4 Hz); 4.36 (dd, 1H,—CHaHb-CH—CHcHd-, J=12.2 Hz and J=3.9 Hz); 5.19 (m, 1H,—CHaHb-CH—CHaHb-); 7.18 (m,1H, —NHCO—). MS (MALDI-TOF): M+1=902 (M+H);M+23=924 (M+Na⁺); M+39=940 (M+K⁺)

Example 6 Preparation du1,2-(ditetradecylthioacetyloxy)-3-palmitoylaminopropane

This compound was synthesized according to the method describedhereinabove (example 4) from 3-(palmitoylamino)propane-1,2-diol (example3) and tetradecylthioacetic acid (example 1).

Yield: 8%

Rf (ethyl acetate/cyclohexane 2:8): 0.33

IR: νNH 3319 cm⁻¹; νCO ester 1735 cm⁻¹; νCO amide 1649 cm⁻¹

MP: 82-83° C.

NMR (¹H, CDCl₃): 0.89 (t, 9H, —CH₃, J=6.4 Hz); 1.26 (multiplet, 68H,—CH₂—); 1.60 (multiplet, 6H, —CH₂—CH₂—S— and —CH₂—CH₂-CONH—); 2.18 (t,2H, —CH₂—CH₂-CONH—, J=6.8 Hz); 2.64 (multiplet, 4H, CH₂—CH₂—S—CH₂—COO—);3.22 (s, 2H, —S—CH₂—COO—); 3.24 (s, 2H, —S—CH₂—COO—); 3.47 (m, 1H,—CONH—CHaHb-CH—CHcHd-); 3.62 (m, 1H, —CONH—CHaHb-CH—CHcHd-); 4.23 (dd,1H, —CHaHb-CH—CHcHd-, J=11.9 Hz and J=5.6 Hz); 4.36 (dd, 1H,—CHaHb-CH—CHcHd-, J=12.2 Hz and J=4 Hz); 5.15 (m, 1H, —CHaHb-CH—CHaHb-);5.85 (m, 1H, —NHCO—). MS (MALDI-TOF): M+1=870 (M+H)

Example 7 Preparation of 1.3-di(oleylamino)propan-2-ol

Oleic acid (5.698 g, 0.020 mol) and 1,3-diaminopropan-2-ol (1 g, 0.011mol) were placed in a flask and heated at 190° C. for 2 hours. Thereaction mixture was cooled to room temperature, then taken up inchloroform and washed with water. The aqueous phase was extracted withchloroform and the organic phases were combined, dried on magnesiumsulfate, filtered and evaporated to dryness to yield an oily blackresidue (6.64 g) which was purified by chromatography on silica gel(eluent: dichloromethane/methanol 99:1). The resulting product was thenwashed with ether and filtered.

Yield: 23%

Rf (dichloromethane/methanol 95:5): 0.43

IR: νNH 3306 cm⁻¹; νCO amide 1646 and 1630 cm⁻¹

MP: 88-92° C.

NMR (¹H, CDCl₃): 0.89 (t, 6H, —CH₃, J=6.2 Hz); 1.28 (multiplet, 68H,—CH₂—); 1.61-1.66 (multiplet, 4H, —CH₂—CH₂-CONH—); 1.98-2.02 (multiplet,8H, —CH₂—CH═CH—CH₂—); 2.23 (t, 4H, —CH₂—CH₂-CONH—, J=7.0 Hz); 3.25-3.42(multiplet, 4H, —CONH—CH₂—CH—CH₂—); 3.73-3.80 (m, 1H,—CONH—CH₂—CH—CH₂—); 5.30-5.41 (multiplet, 4H, —CH₂—CH═CH—CH₂—); 6.36(multiplet, 2H, —NHCO—). MS (MALDI-TOF): M+1=619 (M+H⁺); M+23=641(M+Na⁺); M+39=657 (M+K⁺)

Example 8 Preparation of 1.3-di(tetradecylthioacetylamino)propan-2-ol

This compound was synthesized according to the method describedhereinabove (example 7) from 1,3-diaminopropan-2-ol andtetradecylthioacetic acid (example 1).

Yield: 94%

Rf (dichloromethane/methanol 95:5): 0.44

IR: νNH 3275 cm⁻¹; νCO amide 1660 and 1633 cm⁻¹

MP: 101-104° C.

NMR (1H, CDCl₃): 0.89 (t, 6H, —CH₃, J=6.3 Hz); 1.28 (multiplet, 44H,—CH₂—); 1.57-1.62 (multiplet, 4H, —CH₂—CH₂—S—CH₂—CONH—); 2.55 (t, 4H,—CH₂—CH₂—S—CH₂—CONH—, J=7.2 Hz); 3.26 (s, 4H, —S—CH₂—CONH—); 3.32-3.36(multiplet, 2H, —CONH—CH_(a)H_(b)-CH—CH_(a)H_(b)-NHCO—); 3.43-3.49(multiplet, 2H, —CONH—CH_(a)H_(b)-CH—CH_(a)H_(b)-NHCO—); 3.82-3.84 (m,1H, —CONH—CH₂—CH—CH₂—NHCO—); 7.44 (sl, 2H, —NHCO). MS (MALDI-TOF):M+23=653 (M+Na⁺); M+39=669 (M+K⁺)

Example 9 Preparation of 1,3-di(stearoylamino)propan-2-ol

This compound was synthesized according to the method describedhereinabove (example 7) from 1,3-diaminopropan-2-ol and stearic acid.

Yield: 73%

Rf (dichloromethane/methanol 95:5): 0.28

IR: νNH 3306 cm⁻¹; νCO amide 1647 and 1630 cm⁻¹

MP: 123-130° C.

MS (MALDI-TOF): M+23=645 (M+Na⁺)

Example 10 Preparation of 1.3-diamino-2-(tetradecylthioacetyloxy)propanedihydrochloride Preparation of1.3-di(tert-butyloxycarbonylamino)propan-2-ol (example 10a)

1,3-diaminopropan-2-ol (3 g, 0.033 mol) was dissolved in methanol (300ml) followed by the addition of triethylamine (33 ml dropwise) anddi-tert-butyl dicarbonate [(BOC)₂O] (21.793 g, 0.100 mol). The reactionmedium was heated at 40-50° C. for 20 min then stirred at roomtemperature for 1 hour. After evaporation of the solvent, the colorlessoil residue was purified by chromatography on silica gel (eluent:dichloromethane/methanol 95:5). The reaction yielded a colorless oilwhich crystallized slowly.

Yield: quantitative

Rf (dichloromethane/methanol 95:5): 0.70

IR: νNH 3368 cm⁻¹; νCO carbamate 1690 cm⁻¹

MP: 98-100° C.

NMR (¹H, CDCl₃): 1.45 (multiplet, 18H, —CH₃— (BOC)); 3.02 (sl, 1H, OH);3.15-3.29 (multiplet, 4H, BOCNH—CH₂—CH—CH₂—NHBOC); 3.75 (m, 1H,BOCNH—CH₂—CH—CH₂—NHBOC); 5.16 (multiplet, 2H, —NHBOC). MS (MALDI-TOF):M+1=291 (M+H⁺); M+23=313 (M+Na⁺); M+39=329 (M+K⁺)

Preparation of1.3-di(tert-butyloxycarbonylamino)-2-(tetradecylthioacetyloxy)-propane(example 10b)

1,3-(di-tert-butoxycarbonylamino)-propan-2-ol (example 10a) (1 g, 3.45mmol), tetradecylthioacetic acid (example 1) (0.991 g, 3.45 mmol) anddimethylaminopyridine (0.042 g, 0.34 mmol) were dissolved indichloromethane (40 ml) at 0° C. Dicyclohexylcarbodiimide (0.709 g, 3.45mmol) diluted in dichloromethane was then added dropwise and the mixturewas stirred at 0° C. for 30 min, then brought to room temperature. After20 hours of reaction, the dicyclohexylurea precipitate was filtered andthe filtrate was dried. The oily residue was purified by chromatographyon silica gel (eluent dichloromethane/cyclohexane 5:5 followed bydichloromethane/ethyl acetate 98:2).

Yield: 52%

Rf (dichloromethane/ethyl acetate 95:5): 0.43

IR: νNH 3369 cm⁻¹; νCO carbamate 1690 cm⁻¹; νCO ester 1719 cm⁻¹ NMR (¹H,CDCl₃): 0.89 (t, 3H, CH₃, J=6.3 Hz); 1.26 (multiplet, 22H, —CH₂—); 1.45(multiplet, 18H, —CH₃— (BOC)); 1.56-1.66 (m, 2H, —CH₂—CH₂—S—CH₂—CO);2.64 (t, 2H, —CH₂—CH₂—S—CH₂—CO, J=7.5 Hz); 3.20 (s, 2H, CH₂—S—CH₂—CO);3.35 (multiplet, 4H, BOCNH—CH₂—CH—CH₂—NHBOC); 4.89 (m, 1H,BOCNH—CH₂—CH—CH₂—NHBOC); 5.04 (multiplet, 2H, —NHBOC). MS (MALDI-TOF):M+23=583 (M+Na⁺); M+39=599 (M+K⁺)

Preparation of 1.3-diamino-2-(tetradecylthioacetyloxy)propanedihydrochloride (example 10)

1,3-(ditert-butoxycarbonylamino)-2-tetradecylthioacetyloxypropane(example 10b) (0.800 g, 1.43 mmol) was dissolved in diethyl ether (50ml) saturated with gaseous hydrochloric acid. The reaction medium wasstirred at room temperature for 20 hours. The precipitate which formedwas then filtered and washed with ether.

Yield: 88%

Rf (dichloromethane/methanol 7:3): 0.37

IR: νNH₂ 3049 and 3099 cm⁻¹; νCO ester 1724 cm⁻¹

MP: 224° C. (decomposition)

NMR (¹H, CDCl₃): 0.86 (t, 3H, CH₃, J=6.3 Hz); 1.24 (multiplet, 22H,—CH₂—); 1.48-1.55 (m, 2H, —CH₂—CH₂—S—CH₂—CO); 2.57 (t, 2H,—CH₂—CH₂—S—CH₂—CO, J=7.2 Hz); 3.16 (multiplet, 4H, BOCNH—CH₂—CH—CH₂—NH);3.56 (s, 2H, CH₂—S—CH₂—CO); 5.16 (m, 1H, BOCNH—CH₂—CH—CH₂—NH); 8.43(multiplet, 6H, —NH₂.HCl). MS (MALDI-TOF): M+1=361 (M+H⁺); M+23=383(M+Na⁺); M+39=399 (M+K⁺)

Example 11 Preparation of1,3-ditetradecylthioacetylamino-2-(tetradecylthioacetyloxy)propane

1,3-diamino-2-tetradecylthioacetyloxypropane dihydrochloride (example10) (0.400 g, 0.92 mmol) and tetradecylthioacetic acid (example 1)(0.532 g, 1.84 mmol) were dissolved in dichloromethane (50 ml) at 0° C.followed by the addition of triethylamine (0.3 ml, 2.1 mmol),dicyclohexylcarbodiimide (0.571 g, 2.77 mmol) and hydroxybenzotriazole(HOBt) (0.249 g, 1.84 mmol). The reaction medium was stirred at 0° C.for 1 hour then brought to room temperature for 48 hours. Thedicyclohexylurea precipitate was filtered and washed withdichloromethane. The filtrate was vacuum evaporated. The residueobtained (1.40 g) was purified by chromatography on silica gel (eluent:dichloromethane followed by dichloromethane/ethyl acetate 9:1).

Yield: 74%

Rf (dichloromrthane/ethyl acetate 8:2): 0.25

IR: νNH 3279, 3325 cm⁻¹; νCO ester 1731 cm⁻¹; νCO amide 1647, 1624 cm⁻¹

MP: 87-89° C.

NMR (¹H, CDCl₃): 089 (t, 9H, CH₃, J=6.6 Hz); 1.26 (multiplet, 66H,—CH₂—); 1.55-1.60 (multiplet, 6H, —CH₂—CH₂—S—CH₂—CO); 2.55 (t, 4H,—CH₂—CH₂—S—CH₂—CONH—, J=7.2 Hz); 2.65 (t, 2H, —CH₂—CH₂—S—CH₂—COO—, J=7.2Hz); 3.21 (s, 2H, —CH₂—S—CH₂—COO—); 3.25 (s, 4H, —CH₂—S—CH₂—CONH—);3.40-3.49 (m, 2H, —CONH—CH_(a)H_(b)—CH—CH_(a)H_(b)—NHCO—); 3.52-3.61 (m,2H, —CONH—CH_(a)H_(b)—CH—CH_(a)H_(b)—NHCO—); 4.96 (m,1H,—CONH—CH₂—CH—CH₂—NHCO—); 7.42 (multiplet, 2H, —NHCO—). MS (MALDI-TOF):M+1=901 (M+H⁺); M+23=923 (M+Na⁺); M+39=939 (M+K⁺)

Example 12 Preparation of1.3-dioleylamino-2-(tetradecylthioacetyloxy)propane

This compound was synthesized according to the method described inexample 11 from 1,3-diamino-2-tetradecylthioacetyloxypropanedihydrochloride (example 10) and oleic acid.

Yield: 15%

Rf (dichloromethane/ethyl acetate 8:2): 0.38

IR: νNH 3325 cm⁻¹; νCO ester 1729 cm⁻¹; νCO amide 1640 and 1624 cm⁻¹

MP: 57-59° C.

NMR (¹H, CDCl₃): 0.89 (t, 9H, CH₃, J=6.6 Hz); 1.26 (multiplet, 62H,—CH₂—); 1.59-1.74 (multiplet, 6H, —CH₂—CH₂—S—CH₂—CO); 1.92-2.03(multiplet, 8H, —CH₂—CH═CH—CH₂—); 2.22 (t, 4H, —CH₂—CH₂—S—CH₂—CONH—,J=7.2 Hz); 2.65 (t, 2H, —CH₂—CH₂—S—CH₂—COO—, J=7.4 Hz); 3.19 (s, 2H,—CH₂—S—CH₂—COO—); 3.25-3.34 (m, 2H, —CONH—CHaHb-CH—CHaHb-NHCO—);3.56-3.65 (m, 2H, —CONH—CH_(a)H_(b)—CH—CH_(a)H_(b)—NHCO—); 4.87 (m, 1H,—CONH—CH₂—CH—CH₂—NHCO—); 5.34 (multiplet, 4H, —CH₂—CH═CH—CH₂—); 6.27(multiplet, 2H, —NHCO—). MS (MALDI-TOF): M+1=889 (M+H⁺); M+23=912(M+Na⁺)

Example 13 Preparation of 2.3-ditetradecylthioacetylaminopropan-1-olPreparation of methyl 2.3-diaminopropanoate dihydrochloride (example13a)

2,3-diaminopropionic acid hydrochloride (1 g, 7 mmol) was dissolved inmethanol (40 ml). The medium was cooled in an ice bath, followed by theaddition of thionyl chloride (2.08 ml, 28 mmol). The medium was broughtto room temperature then refluxed for 20 hours. The solvent wasevaporated and the residue was triturated in heptane. The resultingprecipitate was filtered, washed and dried to give a yellowish-whitesolid.

Yield: 94%

Rf: (dichloromethane/methanol 9:1): 0.03

IR: νNH₂ 2811 cm⁻¹; νCO ester 1756 cm⁻¹

MP: 170-180° C. (decomposition)

NMR (¹H, CDCl₃): 3.78 (s, 3H, —CH₃); 4.33 (m, 3H, —CH₂— and —CH—); 8.77(m, 3H, —NH₂.HCl); 9.12 (m, 3H, —NH₂.HCl)

Preparation of methyl 2.3-ditetradecylthioacetylaminopropanoate (example13b)

Methyl 2,3-diaminopropanoate dihydrochloride (example 13a) (0.500 g,2.62 mmol) and tetradecylthioacetic acid (example 1) (1.51 g, 5.23 mmol)were dissolved in dichloromethane (80 ml) at 0° C. followed by theaddition of triethylamine (0.79 ml), dicyclohexylcarbodiimide (1.62 g,7.85 mmol) and hydroxybenzotriazole (0.707 g, 5.23 mmol). The reactionmedium was stirred at 0° C. for 1 hour then brought to room temperaturefor 48 hours. The dicyclohexylurea precipitate was filtered and washedwith dichloromethane and the filtrate was evaporated. The residueobtained (3.68 g) was purified by chromatography on silica gel (eluent:dichloromethane/ethyl acetate 95:5) to give the desired compound in theform of a white powder.

Yield: 96%

Rf: (dichloromethane/methanol 98:2): 0.63

IR: νNH amide 3276 cm⁻¹; νCO ester 1745 cm⁻¹; νCO amide 1649 cm⁻¹

MP: 81.5-82.5° C.

NMR (1H, CDCl₃): 0.89 (t, 6H, CH₃, J=6.6 Hz); 1.26-1.37 (multiplet, 44H,—CH₂—); 1.56-1.61 (m, 4H, —CH₂—CH₂—S—CH₂—CONH); 2.50-2.60 (m, 4H,—CH₂—CH₂—S—CH₂—CONH—); 3.22 (s, 2H, —CH₂—S—CH₂—CONH—); 3.25 (s, 2H,—CH₂—S—CH₂—CONH—); 3.74 (m, 2H, H₃CO(CO)—CH—CH₂—NHCO—); 3.79 (s, 3H,—COOCH₃); 4.64-4.70 (m, 1H, H₃CO(CO)—CH—CH₂—NHCO—); 7.79 (d, 2H, —NHCO—,J=7.3 Hz). MS (MALDI-TOF): M+1=659 (M+H⁺); M+23=681 (M+Na⁺); M+39=697(M+K⁺)

Preparation of 2.3-ditetradecylthioacetylaminopropan-1-ol (example 13)

Sodium borohydride (316 mg, 8.4 mmol) was dissolved in tetrahydrofuran(40 ml). The reaction mixture was cooled in an ice bath followed by theaddition of methyl 2,3-ditetradecylthioacetylaminopropanoate (example13b) (500 mg, 0.76 mmol) in small portions. The mixture was brought toroom temperature and stirred. After 4 days of reaction, 20 ml of waterwere added. The product, which precipitated, was filtered, washed withwater then dried in a dessicator to give a white powder.

Yield: 76%

Rf: (dichloromethane/methanol 95:5): 0.53

IR: νOH alcohol 3436 cm⁻¹; νNH amide 3313 and 3273 cm⁻¹; νCO amide 1648and 1622 cm⁻¹

MP: 100.2-102.2° C.

NMR (¹H, CDCl₃): 0.89 (t, 6H, CH₃, J=6.2 Hz); 1.26 (multiplet, 44H,—CH₂—); 1.59 (m, 4H, —CH₂—CH₂—S—CH₂—CONH); 2.50-2.56 (m, 4H,—CH₂—CH₂—S—CH₂—CONH—); 3.23 (s, 2H, —CH₂—S—CH₂—CONH—); 3.27 (s, 2H,—CH₂—S—CH₂—CONH—); 3.50-3.91 (multiplet, 5H, —OCO—CH₂—CH—CH₂—NHCO—);7.38 (d, 2H, —NHCO—, J=7.1 Hz). MS (MALDI-TOF): M+1=631 (M+H⁺); M+23=653(M+Na⁺); M+39=669 (M+K⁺)

Example 14 Preparation of2.3-ditetradecylthioacetylamino-1-tetradecylthioacetyloxypropane

2,3-ditetradecylthioacetylaminopropan-1-ol (example 13) (0.200 g, 0.32mmol) was dissolved in tetrahydrofuran (40 ml) followed by the additionof dicyclohexylcarbodiimide (65 mg, 0.32 mmol), dimethylaminopyridine(39 mg, 0.32 mmol) and tetradecylthioacetic acid (example 1) (91 mg,0.32 mmol). The mixture was stirred at room temperature for 20 hours.The dicyclohexylurea precipitate was filtered, washed withtetrahydrofuran and the filtrate was evaporated. The residue obtained (1g) was purified by flash chromatography (eluent:dichloromethane) toproduce the desired compound in the form of a white powder.

Yield: 59%

Rf: (dichloromethane/ethyl acetate 8:2): 0.49

IR: νNH amide 3281 cm⁻¹; νCO ester 1736 cm⁻¹; νCO amide 1641 cm⁻¹

MP: 95.4-97.3° C.

NMR (¹H, CDCl₃): 0.89 (t, 9H, CH₃, J=6.4 Hz); 1.27-1.34 (multiplet, 66H,—CH₂—); 1.54-163 (m, 6H, —CH₂—CH₂—S—CH₂—CO—); 2.53 (t, 4H,—CH₂—CH₂—S—CH₂—CONH—, J=7.2 Hz); 2.65 (t, 2H, —CH₂—CH₂—S—CH₂—COO—, J=7.2Hz); 3.21 (s, 2H, —CH₂—S—CH₂—CONH—); 3.23 (s, 2H, —CH₂—S—CH₂—CONH—);3.25 (s, 2H, —CH₂—S—CH₂—COO—); 3.46-3.56 (m, 2H, —OCO—CH₂—CH—CH₂—NHCO—);4.22-4.25 (m, 2H, —OCO—CH₂—CH—CH₂—NHCO—); 4.29-4.39 (m, 1H,—OCO—CH₂—CH—CH₂—NHCO—); 7.29 (t, 1H, —NHCO—); 7.38 (d, 1H, —NHCO—, J=7.6Hz). MS (MALDI-TOF): M+1=901 (M+H⁺)

Example 15 Preparation of1.3-diamino-2-(tetradecylthioacetylthio)propane dihydrochloridePreparation of1,3-di(tert-butyloxycarbonylamino)-2-(p-toluenesulfonyloxy) propane(example 15a)

1,3-di(tert-butyloxycarbonylamino)propan-2-ol (example 10a) (2.89 g, 10mmol) and triethylamine (2.22 ml, 16 mmol) were dissolved in anhydrousdichloromethane (100 ml). The reaction mixture was cooled in an ice bathfollowed by dropwise addition of tosyl chloride (2.272 g, 12 mmol)dissolved in dichloromethane (30 ml). The reaction mixture was thenstirred at room temperature for 72 hours. One equivalent of chloride and1.6 of triethylamine were added after 48 hours. Water was added to stopthe reaction and the medium was allowed to settle. The organic phase waswashed several times with water. The aqueous phases were combined andextracted again with dichloromethane. The organic phase was dried onmagnesium sulfate, filtered and the solvent was evaporated. The residueobtained (6.44 g) was purified by chromatography on silica gel(eluent:dichloromethane followed by dichloromethane/methanol 99:1) toyield the desired compound as a white solid.

Yield: 48%

Rf (dichloromethane/methanol 98:2): 0.70

IR: νNH 3400 cm⁻¹; νCO ester 1716 cm⁻¹; νCO carbamate 1689 cm⁻¹

MP: 104-111° C.

NMR (¹H, CDCl₃):1.42 (s, 18H, CH₃ (BOC)); 2.46 (s, 3H, CH₃); 3.22 and3.41 (multiplet, 4H, BOCNH—CH₂—CH—CH₂—NHBOC); 4.56 (m, 1H,BOCNH—CH₂—CH—CH₂—NHBOC); 5.04-5.11 (multiplet, 2H, —NHBOC); 7.36 (d, 2H,aromatics, J=8.5 Hz); 7.36 (d, 2H, aromatics, J=8.5 Hz). MS (MALDI-TOF):M+23=467 (M+Na⁺); M+39=483 (M+K⁺)

Preparation of 1,3-di(tert-butyloxycarbonylamino)-2-acetylthiopropane(example 15b)

1,3-(ditert-butoxycarbonylamino)-2-(p-toluenesulfonyloxy)propane(example 15a) (0.500 g, 1.12 mmol) and potassium thioacetate (0.161 g,1.41 mmol) were dissolved in acetone and the medium was refluxed for 48hours. One equivalent of potassium thioacetate was added after 24 hoursof reflux. The reaction was brought to room temperature and the solventevaporated. The residue was taken up in diethyl ether and filtered onCelite®. The filtrate was evaporated. The product obtained (0.48 g) waspurified by chromatography on silica gel (eluent: dichloromethane/ethylacetate 98:2) to give the desired compound as an ochre solid.

Yield: 84%

Rf (dichloromethane/methanol 98:2): 0.45

IR: νNH 3350 cm⁻¹; νCO ester 1719 cm⁻¹; νCO carbamate 1691 cm⁻¹

MP: 93-96° C.

NMR (¹H, CDCl₃): 1.45 (s, 18H, CH₃ (BOC)); 2.34 (s, 3H, CH₃); 3.23-3.32(m, 2H, BOCNH—CHaHb-CH—CHaHb-NHBOC); 3.38-3.43 (m, 2H,BOCNH—CHaHb-CH—CHaHb-NHBOC); 3.58-3.66 (m, 1H, BOCNH—CH₂—CH—CH₂—NHBOC);5.22 (multiplet, 2H, —NHBOC). MS (MALDI-TOF): M+23=371 (M+Na⁺)

Preparation of 1,3-di(tert-butyloxycarbonylamino)-2-mercantopropane(example 15c)

1,3-di(tert-butoxycarbonylamino)-2-(acetylthio)propane (example 15b)(0.380 g, 1.2 mmol) diluted in methanol (10 ml) was added to a 20%potassium carbonate solution in methanol (2.14 ml, 12.4 mmol),deoxygenated under a stream of nitrogen. The reaction mixture wasstirred under nitrogen at room temperature for 20 hours, then acidifiedto pH 6 with acetic acid. The solvents were vacuum evaporated. Theresidue was taken up in water and extracted with chloroform. The organicphases were combined, dried on magnesium sulfate, then filtered anddried to give the desired product in the form of a white solid which waspromptly used in the next reaction.

Yield: 90%

Rf (dichloromethane/methanol 98/2): 0.56

IR: νNH 3370 cm⁻¹; νCO carbamate 1680 cm⁻¹

NMR (1H, CDCl₃): 1.46 (s, 18H, CH₃ (BOC)); 2.98-3.12 (multiplet, 3H,BOCNH—CH_(a)H_(b)-CH—CH_(a)H_(b)-NHBOC and BOCNH—CH₂—CH—CH₂—NHBOC);3.46-3.50 (m, 2H, BOCNH—CH_(a)H_(b)-CH—CH_(a)H_(b)-NHBOC); 5.27(multiplet, 2H, —NHBOC).

Preparation of1,3-di(tert-butyloxycarbonylamino)-2-(tetradecylthioacetylthio) propane(example 15d)

1,3-[di(tert-butoxycarbonylamino)]-2-mercaptopropane (example 15c)(0.295 g, 0.963 mmol) was dissolved in dichloromethane (40 ml).Dicyclohexylcarbodiimide (0.199 g, 0.963 mmol), dimethylaminopyridine(0.118 g, 0.963 mmol) and tetradecylthioacetic acid (example 1) (0.278g, 0.963 mmol) were then added. The reaction mixture was stirred at roomtemperature and the progress of the reaction was monitored by TLC. After20 hours of reaction, the dicyclohexylurea precipitate was filtered,washed with dichloromethane and the filtrate was evaporated. The residueobtained (0.73 g) was purified by chromatography on silica gel(eluent:dichloromethane) to give the desired comopund in the form of awhite powder.

Yield: 72%

Rf (dichloromethane/ethyl acetate 95:5): 0.29

IR: νNH 3328 cm⁻¹; νCO thioester 1717 cm⁻¹; νCO carbamate 1687 cm⁻¹

MP: 47-51° C.

NMR (1H, CDCl₃): 0.88 (t, 9H, CH₃, J=6.1 Hz); 1.26 (multiplet, 22H,—CH₂—); 1.44 (s, 18H, CH₃ (BOC)); 1.53-1.65 (m, 2H, —CH₂—CH₂—S—CH₂—CO);2.59 (t, 2H, —CH₂—CH₂—S—CH₂—COS—, J=7.8 Hz); 3.21-3.30 (m, 2H,BOCNH—CH_(a)H_(b)——CH—CH_(a)H_(b)—NHBOC); 3.40 (s, 2H, CH₂—S—CH₂—COS—);3.42-3.49 (m, 2H, BOCNH—CHaHb-CH—CH_(a)H_(b)-NHBOC); 3.62-3.65 (m, 1H,BOCNH—CH₂—CH—CH₂—NHBOC); 5.24 (multiplet, 2H, —NHBOC).

MS (MALDI-TOF): M+23=599 (M+Na⁺); M+39=615 (M+K⁺)

Preparation of 1.3-diamino-2-(tetradecylthioacetylthio)propanedihydrdochloride (example 15)

1,3-[di(tert-butoxycarbonylamino)]-2-tetradecylthioacetylthiopropane(example 15d) (0.300 g, 0.52 mmol) was dissolved in ether saturated ingaseous hydrochloric acid (55 ml). The mixture was stirred at roomtemperature. After 96 hours of reaction, the precipitate which formedwas filtered, washed several times with diethyl ether and dried to givethe desired compound in the form of a white powder.

Yield: 59%

Rf (dichloromethane/methanol 9:1): 0.11

IR: νNH.HCl 2700-3250 cm⁻¹; νCO thioester 1701 cm⁻¹

MP: 181° C. (decomposition)

NMR (1H, CDCl₃): 0.86 (t, 3H, CH₃, J=6 Hz); 1.24 (multiplet, 22H,—CH₂—); 1.49-1.54 (m, 2H, —CH₂—CH₂—S—CH₂—CO); 2.59 (m, 2H,—CH₂—CH₂—S—CH₂—COS—); 2.80-2.84 (m, 1H,BOCNH—CH_(a)H_(b)-CH—CH_(a)H_(b)-NHBOC); 3.03-3.09 (m, 1H,BOCNH—CH_(a)H_(b)-CH—CH_(a)H_(b)-NHBOC); 3.14 (s, 2H, CH₂—S—CH₂—COS—);3.27-3.38 (m, 2H, BOCNH—CH_(a)H_(b)-CH—CH_(a)H_(b)-NHBOC); 3.86-3.90 (m,1H, BOCNH—CH₂—CH—CH₂—NHBOC); 8.21 and 8.52 (2m, 2H+4H, NH₂.HCl).

Example 16 Preparation of1.3-ditetradecylthioacetylamino-2-(tetradecylthioacetylthio)propane

1,3-diamino-2-tetradecylthioacetylthiopropane dihydrochloride (example15) (100 mg, 0.225 mmol) and tetradecylthioacetic acid (example 1) (130mg, 0.450 mmol) were dissolved in dichloromethane (30 ml) at 0° C.followed by the addition of triethylamine (68 μl),dicyclohexylcarbodiimide (139 mg, 0.675 mmol) and hydroxybenzotriazole(61 mg, 0.450 mmol). The reaction mixture was stirred at 0° C. for 1hour then brought to room temperature for 48 hours. The dicyclohexylureaprecipitate was filtered and washed with dichloromethane and thefiltrate was evaporated. The residue obtained (430 mg) was purified bychromatography on silica gel (eluent: dichloromethane/ethyl acetate95:5) to give the desired compound in the form of a white powder.

Yield: 82%

Rf (dichloromethane/methanol 98:2): 0.54

IR: νCO thioester 1660 cm⁻¹; νCO amide 1651 cm⁻¹

MP: 83-85° C.

NMR (¹H, CDCl₃): 0.89 (t, 9H, CH₃, J=6.6 Hz); 1.26 (multiplet, 66H,—CH₂—); 1.56-1.62 (multiplet, 6H, —CH₂—CH₂—S—CH₂—CO); 2.56 (t, 4H,—CH₂—CH₂—S—CH₂—CONH—, J=7.5 Hz); 2.61 (t, 2H, —CH₂—CH₂—S—CH₂—COS—, J=7Hz); 3.26 (s, 4H, CH₂—S—CH₂—CONH—); 3.42 (s, 2H, CH₂—S—CH₂—COS—);3.44-3.49 (m, 2H, —CONH—CH_(a)H_(b)-CH—CH_(a)H_(b)-NH—CO); 3.55-3.61 (m,2H, —CONH—CH_(a)H_(b)-CH—CH_(a)H_(b)-NHCO—); 3.70-3.71 (m,1H,BOCNH—CH₂—CH—CH₂—NHBOC); 7.58-7.62 (m, 2H, NHCO). MS (MALDI-TOF):M+1=917 (M+H⁺); M+23=939 (M+Na⁺)

Example 17 Preparation of1-amino-2.3-di(tetradecylthioacetylthio)propane hydrochloridePreparation of 1-(tert-butyloxycarbonylamino)propane-2.3-diol (example17a)

1-aminopropane-2,3-diol (5 g, 55 mmol) was dissolved in methanol (200ml) followed by dropwise addition of triethylamine (0.5 ml per mmol ofamine) and di-tert-butyl dicarbonate [(BOC)2O] wherein BOC correspondsto tertbutyloxycarbonyl (17.97 g, 82 mmol). The reaction medium washeated at 40-50° C. for 20 min then stirred at room temperature for 1hour. After evaporation of the solvent, the colorless oily residue waspurified by chromatography on silica gel (eluent:dichloromethane/methanol 95:5) to give the desired compound in the formof a colorless oil which crystallized slowly.

Yield: 99%

Rf (dichloromethane/methanol 9:1): 0.39

IR: νNH 3350 cm⁻¹; νCO ester 1746 cm⁻¹; νCO amide 1682 cm⁻¹

MP<15° C.

NMR (1H, CDCl₃):1.44 (s, 9H, CH₃ (BOC)); 3.16-3.31 (m, 2H,BOCNH—CH₂—CH—CH₂—OH); 3.44 (multiplet, 2H, OH); 3.16-3.31 (m, 2H,BOCNH—CH₂—CH—CH₂—OH); 3.71-3.78 (m,1H, BOCNH—CH₂—CH—CH₂—OH); 5.24 (m,1H, —NHBOC). MS (MALDI-TOF): M+23=214 (M+Na⁺)

Preparation of1-(tert-butyloxycarbonylamino)-2.3-di(n-toluenesulfonyloxy) propane(example 17b)

This compound was synthesized according to the method describedhereinabove (example 15a) from1-(tert-butyloxycarbonylamino)-propane-2,3-diol (example 17a) andp-toluenesulfonyl chloride. The reaction produced a white powder.

Yield: 45%

Rf (dichloromethane/methanol 98:2): 0.49

IR: νNH 3430 cm⁻¹; νCO ester and carbamate 1709 cm⁻¹

MP: 112-116° C.

NMR (¹H, CDCl₃): 1.40 (s, 9H, CH₃ (BOC)); 2.46 (s, 6H, CH₃); 3.26-3.45(m, 2H, BOCNH—CH₂—CH—CH₂—OTs); 4.04-4.14 (m, 2H, BOCNH—CH₂—CH—CH₂—OTs);4.68 (m, 1H, BOCNH—CH₂—CH—CH₂—OTs); 4.71 (s, 1H, —NHBOC); 7.34 (d, 4H,aromatics, J=8.5 Hz); 7.69 (d, 2H, aromatics, J=8.1 Hz); 7.76 (d, 2H,aromatics, J=8.1 Hz).

MS (MALDI-TOF): M+23=522 (M+Na⁺); M+39=538 (M+K⁺)

Preparation of 1-(tert-butyloxycarbonylamino)-2.3-di(acetylthio)propane(example 17c)

This compound was synthesized according to the method describedhereinabove (example 15b) from1-(tert-butyloxycarbonylamino)-2,3-di(p-toluenesulfonyloxy)-propane(example 17b) and potassium thioacetate. The reaction produced a whitesolid.

Yield: 59%

Rf (dichloromethane/ethyl acetate 95:5): 0.55

IR: νNH 3430 cm⁻¹; νCO thioester 1718 cm⁻¹; νCO carbamate 1690 cm⁻¹

MP: 62-63° C.

NMR (¹H, CDCl₃): 1.45 (s, 9H, CH₃ (BOC)); 2.35 (s, 3H, CH₃); 2.37 (s,3H, CH₃); 3.12-3.38 (multiplet, 4H, BOCNH—CH₂—CH—CH₂—SCO—); 3.69-3.78(m, 1H, BOCNH—CH₂—CH—CH₂—SCO—); 5.02 (s,1H, —NHBOC). MS (MALDI-TOF):M+23=330 (M+Na⁺)

Preparation of 1-(tert-butyloxycarbonylamino)-2.3-dimercantopropane(example 17d)

This compound was synthesized according to the method describedhereinabove (example 15c) by saponification ofi-(tert-butyloxycarbonylamino)-2,3-di(acetylthio)-propane (example 17c).The reaction produced a white solid which was promptly used in the nextreaction.

Yield: 95%

Rf (dichloromethane/ethyl acetate 95:5): 0.45

IR: νNH 3368 cm⁻¹; νCO carbamate 1688 cm⁻¹

MP: 62-63° C.

NMR (1H, CDCl₃): 1.46 (s, 9H, CH₃ (BOC)); 3.04-3.11 (m, 1H,BOCNH—CH₂—CHSH—CH₂—SH); 3.26-3.35 (m, 2H, BOCNH—CH₂—CHSH—CH₂—SH);3.43-3.52 (m, 2H, BOCNH—CH₂—CH—CH₂—SH); 4.91 (m, 2H, SH); 5.08 (s,1H,—NHBOC).

Preparation of1-(tert-butyloxycarbonylamino)-2.3-di(tetradecylthioacetylthio) pronane(example 17e)

This compound was synthesized according to the method describedhereinabove (example 15d) from1-(tert-butyloxycarbonylamino)-2,3-dimercaptopropane (example 17d) andtetradecylthioacetic acid (example 1). The reaction produced a whitesolid.

Yield: 50%

Rf (dichloromethane): 0.38

IR: νNH 3421 cm⁻¹; νCO thioester 1721 cm⁻¹; νCO carbamate 1683 cm⁻¹

MP: 60-62° C.

NMR (¹H, CDCl₃): 0.87 (t, 6H, CH₃, J=6.3 Hz); 1.26 (multiplet, 44H,—CH₂—); 1.45 (s, 9H, CH₃ (BOC)); 1.57-1.62 (m, 4H, —CH₂—CH₂—S—CH₂—COS—);2.60 (t, 4H, —CH₂—CH₂—S—CH₂—COS—, J=6.9 Hz); 3.17-3.29 (m, 2H,BOCNH—CH_(a)H_(b)-CH—CH_(a)H_(b)—NHBOC); 3.29-3.38 (m, 2H,BOCNH—CH_(a)H_(b)—CH—CH_(a)H_(b)—NHBOC); 3.41 (s, 2H, CH₂—S—CH₂—COS—);3.43 (s, 2H, CH₂—S—CH₂—COS—); 3.76-3.80 (m, 1H, BOCNH—CH₂—CH—CH₂—NHBOC);5.03 (s,1H, —NHBOC). MS (MALDI-TOF): M+23=786 (M+Na⁺)

Preparation of 1-amino-2.3-ditetradecylthioacetylthio)propanehydrochloride (example 17)

This compound was synthesized according to the method describedhereinabove (example 15) from1-(tert-butyloxycarbonylamino)-2,3-ditetradecylthioacetyl-thiopropane(example 17e). The reaction produced a white solid.

Yield: 43%

Rf (dichloromethane): 0.19

IR: νNH.HCl 2700-3250 cm⁻¹; νCO thioester 1701 and 1676 cm⁻¹

MP: 117-128° C.

NMR (¹H, CDCl₃): 0.86 (t, 6H, CH₃, J=6 Hz); 1.24 (multiplet, 44H, —CH₂);1.51 (m, 4H, —CH₂—CH₂—S—CH₂—COS—); 2.61 (m, 4H, —CH₂—CH₂—S—CH₂—COS—);2.93-3.04 (m, 2H, —S—CH_(a)H_(b)—CH—CH_(a)H_(b)—NH₂ HCl); 3.11-3.20 (m,2H, —S—CHaHb-CH—CH_(a)H_(b)—NH₂.HCl); 3.59-3.63 (multiplet, 4H,CH₂—S—CH₂—COS—); 3.72-3.84 (m,1H, —S—CH₂—CH—CH₂—NH₂.HCl); 8.12 (m, 3H,NH₂.HCl).

Example 18 Preparation of1-tetradecylthioacetylamino-2,3-di(tetradecylthioacetylthio)propane

1-amino-2,3-ditetradecylthioacetylthiopropane hydrochloride (example 17)(100 mg, 0.140 mmol) and tetradecylthioacetic acid (example 1) (62 mg,0.210 mmol) were dissolved in dichloromethane (40 ml) at 0° C. followedby the addition of triethylamine (43 ml), dicyclohexylcarbodiimide (59mg, 0.28 mmol) and hydroxybenzotriazole (29 mg, 0.210 mmol). Thereaction mixture was stirred at 0° C. for 1 hour then brought to roomtemperature for 24 hours. It was then heated under gentle reflux for 48hours, then dried. The residue obtained (310 mg) was purified bychromatography on silica gel (eluent: dichloromethane/cyclohexane 8:2)and produced the desired comopund as a white powder.

Yield: 96%

Rf (dichloromethane): 0.20

IR: νNH amide 3306 cm⁻¹; νCO thioester 1674 cm⁻¹; νCO amide 1648 cm⁻¹

MP: 78-80° C.

NMR (¹H, CDCl₃): 0.89 (t, 9H, CH₃, J=6.6 Hz); 1.26 (multiplet, 66H,—CH₂); 1.58-1.62 (multiplet, 6H, —CH₂—CH₂—S—CH₂—COS—); 2.56 (t, 4H,—CH₂—CH₂—S—CH₂—COS—, J=7.5 Hz); 2.61 (t, 2H, —CH₂—CH₂—S—CH₂—CONH—, J=7Hz); 3.26 (s, 4H, CH₂—S—CH₂—COS—); 3.42 (s, 2H, CH₂—S—CH₂—CONH—);3.44-3.49 (m, 2H, —S—CHaHb-CH—CHaHb-NHCO—); 3.55-3.61 (m, 2H,—S—CHaHb-CH—CHaHb-NHCO—); 3.70-3.71 (m, 1H, —S—CH₂—CH—CH₂—NHCO—);7.58-7.62 (m, 1H, NHCO). MS (MALDI-TOF): M+1=934 (M+H⁺); M+23=956(M+Na⁺); M+39=972 (M+K⁺)

Example 19 Preparation of1-tetradecylthioacetylthio-2.3-di(tetradecylthioacetylamino)propanePreparation of 2,3-di(tetradecylthioacetylamino)-1-iodopropane (example19a)

2,3-ditetradecylthioacetylaminopropan-1-ol (example 13) (0.200 g, 0.317mmol) was dissolved in toluene (30 ml). Imidazole (0.054 g, 0.792 mmol),triphenylphosphine (0.208 g, 0.792 mmol) and iodine (0.161 g, 0.634mmol) were then added in that order and the reaction was heated at75-80° C. with stirring. After 6 hours of reaction, the solvent wasevaporated and the residual product was used without furtherpurification. Rf (dichloromethane/methanol 98:2): 0.55

Preparation of 2,3-di(tetradecylthioacetylamino)-1-mercaptopronane(example 19b)

Sodium hydrogen sulfide (0.089 g, 1.59 mmol) was added to2,3-ditetradecylthioacetylamino-1-iodopropane (example 19a) (0.235 g,0.32 mmol) dissolved in acetone (80 ml). The reaction medium was heatedat 70° C. for 16 hours. The solvent was evaporated and the residue takenup in water and extracted with chloroform. The aqueous phase wasacidified to pH 6 with acetic acid, then extracted again withchloroform. The organic phases were dried on magnesium sulfate andfiltered and the solvent was evaporated. The residue obtained was usedwithout further purification.

Preparation of1-tetradecylthioacetylthio-2.3-di(tetradecylthioacetylamino) propane(example 19)

2,3-ditetradecylthioacetylamino-1-mercaptopropane (example 19b) (0.205g, 0.32 mmol) was dissolved in tetrahydrofuran (50 ml).Dicyclohexylcarbodiimide (98 mg, 0.47 mmol), dimethylaminopyridine (58mg, 0.47 mmol) and tetradecylthioacetic acid (example 1) (137 mg, 0.47mmol) were then added. The mixture was stirred at room temperature for20 hours. The dicyclohexylurea precipitate was filtered, washed withtetrahydrofuran and the filtrate was evaporated. The residue obtained(1.14 g) was purified by chromatography on silica gel (eluent:dichloromethane) to give the desired compound in the form of an ochrepowder.

Yield: 10%

Rf (dichloromethane/ethyl acetate 98:2): 0.19

IR: νCO thioester 1711-1745 cm⁻¹; νCO amide 1651 cm⁻¹

MP: 48.8-49.8° C.

NMR (¹H, CDCl₃): 0.89 (t, 9H, CH₃, J=6.3 Hz); 1.26 (multiplet, 66H,—CH₂); 1.58 (m, 6H, —CH₂—CH₂—S—CH₂—COS—); 2.46-55 (m, 4H,—CH₂—CH₂—S—CH₂—CONH); 2.65 (t, 2H, —CH₂—CH₂—S—CH₂—COS—, J=7.4 Hz); 3.24(s, 2H, CH₂—S—CH₂—CONH—); 3.26 (s, 2H, CH₂—S—CH₂—CONH—); 3.66 (t, 2H,—COS—CH₂—CH—CH₂—NHCO); 3.79 (t, 2H, CH₂—S—CH₂—COS—, J=6.3 Hz); 4.31-4.41(m, 2H, —COS—CH₂—CH—CH₂—NHCO); 5.00-5.05 (m, 1H, —COS—CH₂—CH—CH₂—NHCO);7.33 (sl, 1H, NHCO); 9.27 (d, 1H, NHCO, J=8.6 Hz). MS (MALDI-TOF):M+1=917 (M+H⁺); M+23=939 (M+Na⁺); M+39=955 (M+K⁺)

Example 20 Preparation of3-tetradecylthioacetylamino-2-tetradecylthioacetylthiopropan-1-olPreparation of3-tetradecylthioacetylamino-1-trinhenylmethyloxyoroPan-2-ol (example20a)

Chlorotriphenylmethane (2.833 g, 10.16 mmol) was added to a solution of3-tetradecylthioacetylaminopropane-1,2-diol (example 2) (3 g, 8.30 mmol)in pyridine (2.5 ml). The reaction mixture was stirred at 50° C. for 24hours and the solvent was then vacuum evaporated. The residue was takenup in water and extracted with dichloromethane. The organic phase waswashed with 1 N aqueous hydrochloric acid then with an aqueous solutionsaturated in sodium chloride. It was dried on magnesium sulfate,filtered and the solvent was evaporated. The residue obtained (6.36 g)was purified by chromatography on silica gel (eluent:dichloromethane/ethyl acetate 98:2) to give the desired compound in theform of a white powder.

Yield: 69%

Rf (dichloromethane/ethyl acetate 8:2): 0.61

IR: νNH amide 3225 cm⁻¹; νCO amidel654 cm⁻¹

MP: 62.6-65.4° C.

NMR (¹H, CDCl₃): 0.89 (t, 3H, CH₃, J=6.7 Hz); 1.26 (multiplet, 22H,—CH₂); 1.50-1.57 (m, 2H, —CH₂—CH₂—S—CH₂—CONH—); 2.48 (t, 2H,—CH₂—CH₂—S—CH₂—CONH, J=7.2 Hz); 3.01 (m, 1H, OH); 3.17 (s, 2H,CH₂—S—CH₂—CONH—); 3.19 (m, 2H, —O—CH₂—CH—CH₂—NHCO ortrityl-O—CH₂—CH—CH₂—NHCO); 3.27-3.36 (m, 1H, —O—CH₂—CH—CH₂—NHCO ortrityl-O—CH₂—CH—CH₂—NHCO); 3.54-3.62 (m, 1H, —O—CH₂—CH—CH₂—NHCO ortrityl-O—CH₂—CH—CH₂—NHCO); 3.93 (m, 1H, —O—CH₂—CH—CH₂—NHCO); 7.16 (t,1H, NHCO, J=5.7 Hz); 7.23-7.35 (multiplet, 9H, aromatic H); 7.41-7.45(multiplet, 6H, aromatic H). MS (MALDI-TOF): M+23=626 (M+Na⁺).

Preparation of2-iodo-3-tetradecylthioacetylamino-1-triphenylmethyloxypropane (example20b)

3-tetradecylthioacetylamino-1-triphenylmethyloxypropan-2-ol (example20a) (2 g, 3.31 mmol) was dissolved in toluene (100 ml). Imidazole(0.564 g, 8.28 mmol), triphenylphosphine (2.171 g, 8.28 mmol) and iodine(1.681 g, 6.62 mmol) were then added in that order. The reaction mediumwas stirred at room temperature for 20 hours. A saturated sodiumbisulfite solution was added until complete blanching of the reactionmedium. The phases were separated and the aqueous phase was extractedwith toluene. The organic phases were combined, washed with saturatedsodium chloride solution, dried on magnesium sulfate and filtered. Theresidue obtained after evaporation of the solvent (4.65 g) was purifiedby chromatography on silica gel (eluent: dichiromethane) to give thedesired compound in the form of a yellow oil.

Yield: 21%

Rf (dichloromethane/ethyl acetate 95:5): 0.58

IR: νCO amide 1668 cm⁻¹; νCH arom. monosubstituted 748 and 698 cm⁻¹ NMR(¹H, CDCl₃): 0.89 (t, 3H, CH₃, J=6.5 Hz); 1.26 (multiplet, 20H, —CH₂);1.53-1.63 (m, 2H, —CH₂—CH₂—CH₂—S—CH₂—CONH—); 2.63 (m, 2H,—CH₂—CH₂—CH₂—S—CH₂—CONH); 3.13-3.30 (m, 2H, —CH₂—CH₂—S—CH₂—CONH); 3.34(s, 2H, CH₂—S—CH₂—CONH—); 3.67-3.71 (m, 2H, —O—CH₂—CH—CH₂—NHCO ortrityl-O—CH₂—CH—CH₂—NHCO); 3.88-3.94 (m, 2H, —O—CH₂—CH—CH₂—NHCO ortrityl-O—CH₂—CH—CH₂—NHCO); 4.76 (m, 1H, —O—CH₂—CH—CH₂—NHCO); 7.25-7.36(multiplet, 9H, aromatic H); 7.45-7.49 (multiplet, 6H, aromatic H). MS(MALDI-TOF): M-127=586 (M−1)

Preparation of2-mercapto-3-tetradecylthioacetylamino-1-triphenylmethyloxyproPane(example 20c)

Sodium hydrogen sulfate hydrate (38 mg, 0.68 mmol) was prepared as asuspension in ethanol (20 ml) followed by the addition of2-iodo-3-tetradecylthioacetylamino-1-triphenylmethyloxypropane (example20b) (200 mg, 0.28 mmol). The reaction medium was heated at 70° C. 238mg of sodium hydrogen sulfate hydrate were added over several days.After 6.5 days, the solvent was evaporated and the residue taken up indichloromethane and washed with water. The aqueous phase wasre-extracted and the combined organic phases were washed with 0.5Nhydrochloric acid then with saturated sodium chloride solution, thendried on magnesium sulfate. The salt was filtered and the solventevaporated. The residue obtained was used without further purification.

Rf (dichloromethane/ethyl acetate 95:5): 0.33

Preparation of3-tetradecylthioacetylamino-2-tetradecylthioacetylthio-1-triphenylmethyloxy-pronane(example 20d)

2-mercapto-3-tetradecylthioacetylamino-1-triphenylmethyloxypropane(example 20c) (174 mg, 0.28 mmol) was dissolved in tetrahydrofuran (20ml). Dicyclohexylcarbodiimide (88 mg, 0.42 mmol), dimethylaminopyridine(51 mg, 0.42 mmol) and tetradecylthioacetic acid (121 mg, 0.42 mmol)were then added and the reaction medium was stirred at room temperature.After 20 hours of reaction, the solvent was evaporated and the residueobtained (450 mg) was purified by flash chromatography (eluent:dichloromethane/cyclohexane 3:7 to 5-5) to give the desired compound inthe form of a white powder.

Yield: 76%

Rf (dichloromethane): 0.39

IR: νCO thioester and amide 1745 to 1640 cm⁻¹

MP: 48.5-51.9° C.

NMR (1H, CDCl₃): 0.89 (t, 6H, CH₃, J=6.3 Hz); 1.26 (multiplet, 44H,—CH₂); 1.62 (m, 4H, —CH₂—CH₂—S—CH₂—CO—); 2.42 (t, 2H,—CH₂—CH₂—S—CH₂—CONH—, J=7.5 Hz); 2.68 (t, 2H, —CH₂—CH₂—S—CH₂—COS—, J=7.5Hz); 3.14 (s, 2H, CH₂—S—CH₂—CONH—); 3.25 (s, 2H, CH₂—S—CH₂—COS—);3.50-3.59 (m, 1H, —O—CH₂—CH—CH₂—NHCO or trityl-O—CH₂—CH—CH₂—NHCO);3.66-3.72 (m, 1H, —O—CH₂—CH—CH₂—NHCO or trityl-O—CH₂—CH—CH₂—NHCO); 3.96(m, 1H, —O—CH₂—CH—CH₂—NHCO or trityl-O—CH₂—CH—CH₂—NHCO); 3.54-3.62 (m,1H, —O—CH₂—CH—CH₂—NHCO or trityl-O—CH₂—CH—CH₂—NHCO); 5.16 (m, 1H,—O—CH₂—CH—CH₂—NHCO); 7.04 (m, 1H, NHCO, J=5.7 Hz); 7.25-7.34 (multiplet,9H, aromatic H); 7.42-7.45 (multiplet, 9H, aromatic H).

MS (MALDI-TOF): M+23=889 (M+Na⁺)

Preparation of3-tetradecylthioacetylamino-2-tetradecylthioacetylthiopropan-1-ol(example 20)

3-tetradecylthioacetylamino-2-tetradecylthioacetylthio-1-triphenylmethyloxy-propane(example 20d) (187 mg, 0.21 mmol) was dissolved in ether saturated withgaseous hydrochloric acid (12 ml). The reaction medium was stirred atroom temperature for 20 hours. The precipitate which formed was filteredand washed with diethyl ether to give the desired compound in the formof a white powder.

Yield: 52%

Rf (dichloromethane/methanol 98:2): 0.48

IR: νCO thioester 1704 cm⁻¹; νCO amide 1646 cm⁻¹

MP: 88.4-94.1° C.

NMR (1H, CDCl₃): 0.89 (t, 6H, CH₃, J=6.4 Hz); 1.26-1.37 (multiplet, 44H,—CH₂); 1.55-1.61 (m, 4H, —CH₂—CH₂—S—CH₂—CO—); 2.55 (t, 2H,—CH₂—CH₂—S—CH₂—CONH—, J=7 Hz); 2.65 (t, 2H, —CH₂—CH₂—S—CH₂—COS—, J=7Hz); 3.26 (s, 2H, CH₂—S—CH₂—CONH—); 3.27 (s, 2H, CH₂—S—CH₂—COS—);3.36-3.38 (m, 1H, —O—CH₂—CH—CH₂—NHCO); 3.58-3.64 (m, 1H,—O—CH₂—CH—CH₂—NHCO); 4.02 (m, 1H, —O—CH₂—CH—CH₂—NHCO); 4.11-4.25 (m, 2H,HO—CH₂—CH—CH₂—NHCO); 7.34 (m,1H, NHCO).

MS (MALDI-TOF): M+23=670 (M+Na⁺)

Example 21 Preparation of3-tetradecylthioacetylamino-1-tetradecylthiacetyloxy-2-tetradecylthioacetylthiopropane

3-tetradecylthioacetylamino-2-tetradecylthioacetylthiopropan-1-ol(example 20) (64 mg, 0.10 mmol) was dissolved in tetrahydrofuran (7 ml).Dicyclohexylcarbodiimide (31 mg, 0.15 mmol), dimethylaminopyridine (18mg, 0.15 mmol) and tetradecylthioacetic acid (example 1) (43 mg, 0.15mmol) were then added. The mixture was stirred at room temperature for20 hours. The dicyclohexylurea precipitate was filtered and the filtratewas evaporated. The residue obtained (140 mg) was purified by flashchromatography (eluent: dichloromethane) to give the desired compound inthe form of a white powder.

Yield: 17%

Rf (dichloromethane/ethyl acetate 98:2): 0.23

IR: νCO ester 1730 cm⁻¹; νCO thioester 1671 cm⁻¹; νCO amide 1645 cm⁻¹

MP: 59.0-63.4° C.

NMR (1H, CDCl₃): 0.89 (t, 9H, CH₃, J=6.5 Hz); 1.26-1.37 (multiplet, 66H,—CH₂); 1.58-1.63 (m, 6H, —CH₂—CH₂—S—CH₂—CO—); 2.53 (t, 2H,—CH₂—CH₂—S—CH₂—CONH—, J=7.6 Hz); 2.61-2.67 (m, 4H, —CH₂—CH₂—S—CH₂—COS—and —CH₂—CH₂—S—CH₂—COO); 3.23 (s, 4H, CH₂—S—CH₂—CONH— andCH₂—S—CH₂—COO—); 3.24 (s, 2H, CH₂—S—CH₂—COS—); 3.50-3.57 (m, 1H,—O—CH₂—CH—CH₂—NHCO); 3.63-3.72 (m, 1H, —O—CH₂—CH—CH₂—NHCO); 4.19-4.25(m, 1H, —O—CH₂—CH—CH₂—OCO); 3.63-3.72 (m, 1H, —O—CH₂—CH—CH₂—OCO); 5.19(m, 1H, —O—CH₂—CH—CH₂—NHCO); 7.20 (m, 1H, NHCO).

MS (MALDI-TOF): M+23=940 (M+Na⁺)

Example 22 Preparation of1-amino-2-tetradecylthioacetyloxy-3-tetradecylthioacetylthiopropanehydrochloride Preparation of1-tert-butyloxycarbonylamino-3-iodopronan-2-ol (example 22a)

1-[(tert-butyloxycarbonyl)amino]propane-2,3-diol (example 17a) (3.88 g,20 mmol) was dissolved in toluene (250 ml). Imidazole (1.73 g, 25 mmol),triphenylphosphine (6.65 g, 25 mmol) and iodine (5.15 g, 20 mmol) werethen added in that order. The reaction medium was stirred at roomtemperature for 17 hours and 0.5 equivalents of imidazole,triphenylphosphine and iodine were added. After 21 hours of reaction, asaturated sodium sulfite solution was added until complete blanching ofthe reaction medium. The phases were allowed to settle and the aqueousphase was extracted twice with toluene. The combined organic phases werewashed with saturated sodium chloride solution, dried on magnesiumsulfate, filtered and the solvent evaporated. The residue obtained(11.02 g) was purified by chromatography on silica gel (eluent:dichloromethane/ethyl acetate 95:5) to give the desired compound as ayellow paste which was promptly used in the next reaction.

Yield: 41%

Rf (dichloromethane/methanol 98:2): 0.24

IR: νNH amide 3387 cm⁻¹; νCO carbamate 1678 cm⁻¹

Prenaration of 3-acetylthio-1-tert-butyloxycarbonylaminopropan-2-ol(examle 22b)

1-(tert-butyloxycarbonylamino)-3-iodopropan-2-ol (example 22a) (2 g,6.64 mmol) and potassium thioacetate (0.948 g, 8.30 mmol) were dissolvedin acetone (30 ml) and the medium was refluxed for 16 hours. The solventwas vacuum evaporated and the residue was taken up in diethyl ether,then filtered on Celite®. The filtrate was evaporated. The residueobtained (1.69 g) was purified by chromatography on silica gel (eluent:dichloromethane/ethyl acetate 98:2) then repurified by flashchromatography (eluent: dichloromethane) to give the desired compound inthe form of a yellow oil.

Yield: 27%

Rf (dichloromethane/ethyl acetate 95:5): 0.31

IR: νNH amide 3367 cm⁻¹; νCO thioester 1744 cm⁻¹; νCO carbamate 1697cm⁻¹ NMR (1H, CDCl₃): 1.26 (m, 9H, CH₃ (boc)); 2.37 (s, 3H, COCH₃); 3.04(m, 1H, —NH—CH₂—CH—CH₂—S— or —NHCH₂—CH—CH₂—S—); 3.24 (m, 1H,—NH—CH₂—CH—CH₂—S-ou —NHCH₂—CH—CH₂—S—); 3.30-3.41 (m, 2H,—NH—CH₂—CH—CH₂—S— or —NHCH₂—CH—CH₂—S—); 4.86 (sl, 1H, OH); 4.96 (m, 1H,—NH—CH₂—CH—CH₂—S—).

Preparation of 1-tert-butyloxycarbonylamino-3-mercaptopronan-2-ol(example 22c)

3-acetylthio-1-tert-butyloxycarbonylaminopropan-2-ol (example 22b)(0.307 g, 1.23 mmol) diluted in a minimum of methanol (7 ml) was addedto a 20% potassium carbonate solution (3.49 ml, 12.31 mmol) in methanol,deoxygenated under a stream of nitrogen. The medium was stirred at roomtemperature under a stream of nitrogen for 20 hours, then acidified topH 6 with acetic acid and concentrated to dryness. The residue obtainedwas taken up in water and extracted with dichloromethane. The organicphase was dried on magnesium sulfate, filtered and concentrated. Theoily residue obtained was used immediately in the next reaction withoutfurther purification.

Yield: 78%

Rf (dichloromethane/ethyl acetate): 0.07

Preparation of1-tert-butyloxycarbonylamino-2-tetradecylthioacetyloxy-3-tetradecylthioacetylthiopronane(example 22d)

1-(tert-butyloxycarbonylamino)-3-mercaptopropan-2-ol (example 22c)(0.200 g, 96 mmol) was dissolved in dichloromethane (50 ml).Dicyclohexylcarbodiimide (0.398 g, 1.93 mmol), dimethylaminopyridine(0.236 g, 1.93 mmol) and tetradecylthioacetic acid (example 1) (0.557 g,1.93 mmol) were then added. The mixture was stirred at room temperaturefor 20 hours. The dicyclohexylurea precipitate was filtered, washed withdichloromethane and the filtrate was evaporated. The residue obtained(1.2 g) was purified by chromatography on silica gel (eluent:dichloromethane) to give the desired compound in the form of a whitepaste.

Yield: 47%

Rf (dichloromethane): 0.26

IR: νNH amide 3314 cm⁻¹; νCO ester, amide and thioester 1682 to 1744cm⁻¹ NMR (1H, CDCl₃): 0.89 (t, 6H, CH₃, J=6.5 Hz); 1.27 (multiplet, 40H,CH₂); 1.45 (multiplet, 9H, CH₃ (BOC)); 1.56-1.63 (m, 4H,—CH₂—CH₂—CH₂—S—CH₂—CO—); 2.65 (m, 4H, —CH₂—CH₂—S—CH₂—CO—); 2.92 (s, 4H,—CH₂—S—CH₂—CO—); 2.96 (m, 4H, —CH₂—S—CH₂—CO—); 3.24-3.40 (m, 2H,—NH—CH₂—CH—CH₂—S— or —NHCH₂—CH—CH₂—S); 3.44-3.51 (m, 2H,—NH—CH₂—CH—CH₂—S— or —NHCH₂—CH—CH₂—S—); 4.91 (m, 1H, —NH—CH₂—CH—CH₂—S—);5.19 (m, 1H, NHCO).

MS (MALDI-TOF): M+23=770 (M+Na⁺)

Preparation of1-amino-2-tetradecylthioacetyloxy-3-tetradecylthioacetylthio propanehydrochloride (example 22)

1-(tert-butoxycarbonylamino)-2-tetradecylthioacetyloxy-3-tetradecylthioacetyl-thiopropane(example 22d) (300 mg, 0.40 mmol) was dissolved in diethyl ethersaturated with gaseous hydrochloric acid (70 ml) and the reaction mediumwas stirred at room temperature for 72 hours. The precipitate whichformed was filtered, washed with diethyl ether and dried to give thedesired compound in the form of a white powder.

Yield: 42%

IR: νCO ester 1733 cm⁻¹; νCO thioester 1692 cm⁻¹

MP: 82° C. (decomposition) NMR (1H, CDCl₃): 0.86 (t, 6H, CH₃, J=6.6 Hz);1.24 (multiplet, 44H, —CH₂); 1.52 (m, 4H, —CH₂—CH₂—S—CH₂—CO—); 2.52-2.62(m, 4H, —CH₂—CH₂—S—CH₂—CO—); 3.07-3.15 (multiplet, 4H,—S—CH₂—CH—CH₂—NH₂); 3.40 (s, 2H, CH₂—S—CH₂—COO—); 3.61 (s, 2H,CH₂—S—CH₂—COS—); 5.12 (m, 1H, —S—CH₂—CH—CH₂—NH₂); 8.01 (m, 3H, —NH₂.HCl)

Example 23 Preparation of1-tetradecylthioacetylamino-2-tetradecylthiacetyloxy-3-tetradecylthioacetylthioproPane

3-amino-2-tetradecylthioacetyloxy-1-tetradecylthioacetyl-thiopropanehydrochloride (example 22) (100 mg, 0.15 mmol) and tetradecylthioaceticacid (example 1) (63 mg, 0.22 mmol) were dissolved in dichloromethane(30 ml) at 0° C. followed by the addition of triethylamine (0.044 ml),dicyclohexylcarbodiimide (60 mg, 0.29 mmol) and hydroxybenzotriazole (30mg, 0.22 mmol). The reaction medium was stirred at 0° C. for 1 hour thenbrought to room temperature for 48 hours. The dicyclohexylureaprecipitate was filtered, washed with dichloromethane and the filtratewas evaporated. The residue obtained (263 mg) was purified by flashchromatography (eluent: dichloromethane/ethyl acetate 98:2) to give thedesired compound in the form of a white powder.

Yield: 98%

Rf (dichloromethane/ethyl acetate 95:5): 0.38

IR: νNH amide 3340 cm⁻¹; νCO ester 1727 cm⁻¹; νCO amide and thioester1655 and 1669 cm⁻¹

MP: 63.9-67.1° C.

NMR (1H, CDCl₃): 0.89 (t, 9H, CH₃, J=6.2 Hz); 1.26 (multiplet, 66H,—CH₂); 1.54-1.66 (m, 6H, —CH₂—CH₂—S—CH₂—CO—); 2.52-2.67 (m, 6H,—CH₂—CH₂—S—CH₂—CO—); 3.08 (m, 1H, —S—CH₂—CH—CH₂—NHCO or—S—CH₂—CH—CH₂—NHCO); 3.21 (s, 2H, CH₂—S—CH₂—CONH—); 3.23 (s, 2H,CH₂—S—CH₂—COO—); 3.27 (m, 1H, —S—CH₂—CH—CH₂—NHCO or —S—CH₂—CH—CH₂—NHCO);3.43 (s, 2H, CH₂—S—CH₂—COS—); 3.50 (m, 1H, —S—CH₂—CH—CH₂—NHCO or—S—CH₂—CH—CH₂—NHCO); 3.62 (m, 1H, —S—CH₂—CH—CH₂—NHCO or—S—CH₂—CH—CH₂—NHCO); 5.06 (m, 1H, —COS—CH₂—CH—CH₂—NHCO); 7.24 (t,1H,—NHCO, J=6.7 Hz)

MS (MALDI-TOF): M+1=918 (M+H⁺); M+23=940 (M+Na⁺)

Example 24 Method of Preparation of the Compounds Represented by Formula(I) According to the Invention.

To perform the in vitro experiments described in the following examples,the inventive compounds were prepared in the form of an emulsion asdescribed below.

An emulsion comprising an inventive compound and phosphatidylcholine(PC) was prepared as described by Spooner et al. (Spooner, Clark et al.1988). The inventive compound was mixed with PC in a 4:1 (m/m) ratio inchloroform, the mixture was dried under nitrogen, then vacuum evaporatedovernight; the resulting powder was taken up in 0.16 M KCl containing0.01 M EDTA and the lipid particles were then dispersed by ultrasoundfor 30 minutes at 37° C. The liposomes so formed were then separated byultracentrifugation (XL 80 ultracentrifuge, Beckman Coulter, Villepinte,France) at 25,000 rpm for 45 minutes to recover liposomes having a sizegreater than 100 nm and close to that of chylomicrons. Liposomescomposed only of PC were prepared concurrently to use as negativecontrol.

The composition of the liposomes in the inventive compound was estimatedby using the enzyme calorimetric triglyceride assay kit. The assay wascarried out against a standard curve, prepared with the lipid calibratorCFAS, Ref. 759350 (Boehringer Mannheim GmbH, Germany). The standardcurve covered concentrations ranging from 16 to 500 μg/ml. 100 μl ofeach sample dilution or calibration standard were deposited per well ona titration plate (96 wells). 200 μl of triglyceride reagents, ref.701912 (Boehringer Mannheim GmbH, Germany) were then added to each well,and the entire plate was incubated at 37° C. for 30 minutes. Opticaldensities (OD) were read on a spectrophotometer at 492 nm. Triglycerideconcentrations in each sample were calculated from the standard curveplotted as a linear function y=ax+b, where y represents OD and xrepresents triglyceride concentrations.

Liposomes containing the inventive compounds, prepared in this manner,were used for in vitro experiments described in examples 26, 27 and 28.

Example 25 Evaluation of the Antioxidant Properties of the InventiveCompounds

A Protection against LDL oxidation induced by copper: Oxidation of LDLis an important modification which plays a major role in the onset anddevelopment of atherosclerosis (Jurgens, Hoff et al. 1987). Thefollowing protocol allows demonstration of the antioxidant properties ofcompounds. Unless otherwise indicated, all reagents were from Sigma (StQuentin, France).

LDL were prepared as described in Lebeau et al. (Lebeau, Furman et al.2000). The solutions of the test compounds were prepared at 10⁻² M inethanol and diluted in PBS so that the final concentration ranged from0.1 to 100 μM with a total ethanol concentration of 1% (VN).

Before oxidation, EDTA was removed from the LDL preparation by dialysis.The oxidation reaction was then carried out at 30° C. by adding 100 μlof 16.6 μM CuSO₄ to 800 μl of LDL (125 μg protein/ml) and 100 μl of atest compound solution. The formation of dienes, the species to befollowed, was measured by the optical density at 234 nm in the samplestreated with the compounds in the presence or absence of copper. Opticaldensity at 234 nm was measured every 10 minutes for 8 hours on athermostated spectrophotometer (Kontron Uvikon 930). The analyses werecarried out in triplicate. A compound was considered to have antioxidantactivity when it shifted the lag phase latency relative to the controlsample. The inventors demonstrate that the inventive compounds delayedLDL oxidation (induced by copper), indicating that the inventivecompounds possess intrinsic antioxidant activity. FIG. 2 presents anexample of the results obtained with the inventive compounds.

FIG. 2 shows that inventive compounds Ex 2, 4, 5, 6 et 11, exhibitintrinsic antioxidant properties.

FIG. 2 a shows that the inventive compounds shifted the lag phaselatency by more than 13% for compound Ex 2 up to 34.3% for compound Ex4. The inventive compounds did not appear to modify the oxidation rate(FIG. 2 b) or the amount of dienes formed (FIG. 2 c).

B—Evaluation of the protection conferred by the inventive compoundsagainst lipid peroxidation:

The inventive compounds which were tested are the compounds whosepreparation is described in examples 2 to 23.

LDL oxidation was measured by the TBARS method (Thiobarbituric AcidReactive Substrates).

According to the same principle as that described hereinabove, LDL wereoxidized in the presence of CuSO₄ and lipid peroxidation was evaluatedas follows:

TBARS were measured by a spectrophotometric method, lipidhydroperoxidation was measured by using lipid peroxide-dependentoxidation of iodide to iodine.

The results are expressed as nmol of malondialdehyde (MDA) or as nmolhydroperoxide/mg protein.

The results obtained hereinabove by measuring the inhibition ofconjugated diene formation, were confirmed by the experiments measuringLDL lipid peroxidation. Thus, the inventive compounds also affordedefficient protection of LDL against lipid peroxidation induced by copper(an oxidizing agent).

Example 26 Measurement of the Antioxidant Properties of the InventiveCompounds on Cell Cultures

A—Culture Protocol:

Neuronal, neuroblastoma (human) and PC12 cells (rat) were the cell linesused for this type of study. PC12 cells were prepared from a ratpheochromocytoma and have been characterized by Greene and Tischler(Greene and Tischler, 1976). These cells are commonly used in studies ofneuron differentiation, signal transduction and neuronal death. PC12cells were grown as previously described (Farinelli, Park et al. 1996)in complete RPMI medium (Invitrogen) supplemented with 10% horse serumand 5% fetal calf serum. Primary cultures of endothelial and smoothmuscle cells were also used. Cells were obtained from Promocell(Promocell GmBH, Heidelberg) and cultured according to the supplier'sinstructions.

The cells were treated with different doses of the compounds rangingfrom 5 to 100 μM for 24 hours. The cells were then recovered and theincrease in expression of the target genes was evaluated by quantitativePCR.

B—mRNA Measurement:

mRNA was extracted from the cultured cells treated or not with theinventive compounds. Extraction was carried out with the reagents of theAbsolutely RNA RT-PCR miniprep kit (Stratagene, France) as directed bythe supplier. mRNA was then assayed by spectrometry and quantified byquantitative RT-PCR with a Light Cycler Fast Start DNA Master Sybr GreenI kit (Roche) on a Light Cycler System (Roche, France). Primer pairsspecific for the genes encoding the antioxidant enzymes superoxidedismutase (SOD), catalase and glutathione peroxidase (GPx) were used asprobes. Primer pairs specific for the □-actin and cyclophilin genes wereused as control probes.

An increase in mRNA expression of the antioxidant enzyme genes, measuredby quantitative RT-PCR, was demonstrated in the different cell typesused, when the cells were treated with the inventive compounds.

C—Control of Oxidative Stress:

Measurement of Oxidizing Species in the Cultured Cells:

The antioxidant properties of the compounds were also evaluated by meansof a fluorescent tag the oxidation of which is followed by appearance ofa fluorescence signal. The reduction in the intensity of the emittedfluorescence signal was determined in cells treated with the compoundsin the following manner: PC12 cells cultured as described earlier (black96-well plates, transparent bottom, Falcon) were incubated withincreasing doses of H₂O₂ (0.25 mM-1 mM) in serum-free medium for 2 and24 hours. After incubation, the medium was removed and the cells wereincubated with 10 μM dichlorodihydrofluorescein diacetate solution(DCFDA, Molecular Probes, Eugene, USA) in PBS for 30 min at 37° C. in a5% CO₂ atmosphere. The cells were then rinsed with PBS. The fluorescenceemitted by the oxidation tag was measured on a fluorimeter (Tecan Ultra384) at an excitation wavelength of 495 nm and an emission wavelength of535 nm. The results are expressed as the percentage of protectionrelative to the oxidized control. Fluorescence intensity was lower inthe cells incubated with the inventive compounds than in untreatedcells. These findings indicate that the inventive compounds promoteinhibition of the production of oxidative species in cells subjected tooxidative stress. The previously described antioxidant properties arealso effective at inducing antiradical protection in cultured cells.

D—Measurement of Lipid Peroxidation:

The different cell lines (cell models noted hereinabove) and the primarycell cultures were treated as described earlier. The cell supernatantwas recovered after treatment and the cells were lysed and recovered fordetermination of protein concentration. Lipid peroxidation was detectedas follows: lipid peroxidation was measured by using thiobarbituric acid(TBA) which reacts with lipid peroxidation of aldehydes such asmalondialdehyde (MDA). After treatment, the cell supernatant wascollected (900 μl) and 90 μl of butylated hydroxytoluene were added(Morliere, Moysan et al. 1991). One milliliter of 0.375% TBA solution in0.25 M hydrochloric acid containing 15% trichloroacetic acid was alsoadded to the reaction medium. The mixture was heated at 80° C. for 15min, cooled on ice and the organic phase was extracted with butanol. Theorganic phase was analyzed by spectrofluorimetry (λexc=515 nm andλem=550 nm) on a Shimazu 1501 spectrofluorimeter (Shimadzu Corporation,Kyoto, Japan). TBARS are expressed as MDA equivalents usingtetra-ethoxypropane as standard. The results were normalized for proteinconcentration.

The decrease in lipid peroxidation observed in the cells treated withthe inventive compounds confirms the previous results.

The inventive compounds advantageously exhibit intrinsic antioxidantproperties allowing to slow and/or inhibit the effects of an oxidativestress. The inventors also show that the inventive compounds are capableof inducing the expression of genes encoding antioxidant enzymes. Theseparticular features of the inventive compounds allow cells to moreeffectively fight against oxidative stress and therefore be protectedagainst free radical-induced damage.

Example 27 Evaluation of PPAR Activation In Vitro by the InventiveCompounds

Nuclear receptors of the PPAR subfamily which are activated by two majorpharmaceutical classes—fibrates and glitazones, widely used in theclinic for the treatment of dyslipidemias and diabetes—play an importantrole in lipid and glucose homeostasis. The following experimental datashow that the inventive compounds activate PPARa in vitro.

PPAR activation was tested in vitro in RK13 fibroblast cell lines or ina hematocyte line HepG2 by measuring the transcriptional activity ofchimeras composed of the DNA binding domain of the yeast gal4transcription factor and the ligand binding domain of the differentPPARs. The example below is given for HepG2 cells.

A—Culture Protocols:

HepG2 cells were from ECACC (Porton Down, UK) and were grown in DMEMmedium supplemented with 10% (VN) fetal calf serum, 100 U/ml penicillin(Gibco, Paisley, UK) and 2 mM L-glutamine (Gibco, Paisley, UK). Theculture medium was changed every two days. Cells were kept at 37° C. ina humidified 95% air/5% CO₂ atmosphere.

B—Description of Plasmids used for Transfection:

The plasmids pG5TkpGL3, pRL-CMV, pGal4-hPPAR□, pGal4-hPPAR□ and pGal4-fhave been described by Raspe et al. (Raspe, Madsen et al. 1999). ThepGal4-mPPARα and pGal4-hPPARβ constructs were obtained by cloningPCR-amplified DNA fragments corresponding to the DEF domains of themouse PPARα and human PPARα nuclear receptors, respectively, into thepGal4-f vector.

C—Transfection

HepG2 cells were seeded in 24-well culture dishes at 5×10⁴ cells/welland transfected for 2 hours with the reporter plasmid pG5TkpGL3 (50ng/well), the expression vectors pGal4-f, pGal4-mPPARα, pGal4-hPPARα,pGal4-hPPARγ, pGal4-hPPARβ (100 ng/well) and the transfection efficiencycontrol vector pRL-CMV (1 ng/well) according to the previously describedprotocol (Raspe, Madsen et al. 1999), then incubated for 36 hours withthe test compounds. At the end of the experiment, the cells were lysed(Gibco, Paisley, UK) and luciferase activity was determined with aDual-Luciferase™ Reporter Assay System kit (Promega, Madison, Wis., USA)according to the supplier's instructions. The protein content of thecell extracts was then measured with the Bio-Rad Protein Assay kit(Bio-Rad, Munich, Germany) as directed by the supplier.

The inventors demonstrate an increase in luciferase activity in cellstreated with the inventive compounds and transfected with thepGal4-hPPARa plasmid. Said induction of luciferase activity indicatesthat the inventive compounds are activators of PPARA. FIG. 3 gives anexample of the results obtained with the inventive compounds.

FIG. 3: HepG2 cells transfected with Gal4/PPARα plasmids were incubatedwith different concentrations (5, 15, 50 and 100 μM) of the inventivecomopunds (Ex 2, Ex 4, Ex 5, Ex 6, Ex 11) for 24 h and with differentconcentrations of the vehicle (PC) noted 1, 2, 3, 4 as controls for the5, 15, 50 and 100 pM concentrations of the inventive compounds(according to the 4:1 (m/m) ratio described in example 24 (Method ofpreparation of compounds represented by formula (I) according to theinvention)). The results are expressed as the induction factor(luminescent signal of treated cells divided by luminescent signal ofuntreated cells) after the different treatments. The higher theinduction factor the more potent the PPARa agonist activity. The resultsshow that inventive compound Ex 2 produced a maximum 19.8-fold inductionof the luminescent signal at 50 μM, 19.2 at 100 μM, 7.7 at 15 μM and 1.5at 5 μM. Inventive compound Ex 5 also showed a dose-dependent increasein the induction factor of 10.5 at 100 μM, 7 at 50 μM, 2.5 at 15 μM and1.2 at 5 μM. Inventive compound Ex 6 also induced an increase in theluminescent signal, revealing an activity on the PPARa nuclear receptor.The induction factors for inventive compound Ex 6 were 14.5 at 100 μM,9.6 at 50 μM, 2.2 at 15 μM and 1.1 at 5 μM. In contrast, when the cellswere incubated with the vehicle (PC liposome), no significant inductionwas observed. These results demonstrate that the inventive compoundstested exhibit significant PPARa ligand activity and therefore enablethe transcriptional activation thereof.

Example 28 Evaluation of the Anti-Inflammatory Properties of theInventive Compounds

An inflammatory response is observed in many neurological disorders,such as cerebral ischemias. Inflammation is also an important factor inneurodegeneration. In stroke, one of the first reactions of glial cellsis to release cytokines and free radicals. This release of cytokines andfree radicals results in an inflammatory response in the brain which canlead to neuronal death (Rothwell 1997).

Cell lines and primary cells were cultured as described hereinabove.

Lipopolysaccharide (LPS) bacterial endotoxin (Escherichia coli 0111:B4)(Sigma, France) was reconstituted in distilled water and stored at 4° C.Cells were treated with LPS 1 μg/ml for 24 hours. To avoid interferencefrom other factors, the culture medium was completely changed.

TNF-α is an important factor in the inflammatory response to stress(oxidative stress for example). To evaluate TNF-α secretion in responseto stimulation by increasing doses of LPS, the culture medium ofstimulated cells was removed and TNF-α was assayed with an ELISA-TNF-αkit (Immunotech, France). Samples were diluted 50-fold so as to be inthe range of the standard curve (Chang, Hudson et al. 2000).

The anti-inflammatory property of the compounds was characterized asfollows: the cell culture medium was completely changed and the cellswere incubated with the test compounds for 2 hours, after which LPS wasadded to the culture medium at 1 μg/ml final concentration. After a24-hour incubation, the cell supernatant was recovered and stored at−80° C. when not treated directly. Cells were lysed and protein wasquantified with the Bio-Rad Protein Assay kit (Bio-Rad, Munich, Germany)according to the supplier's instructions.

The measurement of the decrease in TNF-α secretion induced by treatmentwith the test compounds is expressed as pg/ml/μg protein and as thepercentage relative to the control. These results show that theinventive compounds have anti-inflammatory properties.

Example 29 Evaluation of the Neuroprotective Effects of the InventiveCompounds in a Cerebral Ischemia-Reperfusion Model

A—Prophylactic Model:

1—Treatment of Animals

1.1 Animals and Administration of the Compounds

Wistar rats weighing 200 to 350 g were used for this experiment.

Animals were maintained on a 12 hour light-dark cycle at a temperatureof 20° C.+3° C. Water and food were available ad libitum. Food intakeand weight gain were recorded.

Animals were treated by gavage with the inventive compounds (600mg/kg/day) in suspension in the vehicle (0.5% carboxycellulose (CMC) and0.1% Tween) or with the vehicle alone, for 14 days before ischemiainduction by occlusion of the middle cerebral artery.

The carboxymethylcellulose used is a sodium salt of intermediateviscosity carboxymethylcellulose (Ref. C4888, Sigma-Aldrich, France).Tween used is Polyoxyethylenesorbitan Monooleate (Tween 80, Ref. P8074,Sigma-Aldrich, France).

1.2 Ischemia Induction-Reperfusion by Intraluminal Occlusion of theMiddle Cerebral Artery:

Animals were anesthetized by intraperitoneal injection of 300 mg/kgchloral hydrate. A rectal probe was inserted and body temperature wasmaintained at 37° C.±0.5° C. Blood pressure was monitored throughout theexperiment. Under a surgical microscope, the right carotid artery wasexposed by a median incision in the neck. The pterygopalatine artery wasligated at its origin and an arteriotomy was fashioned in the externalcarotid artery so as to insert a nylon monofilament, which was gentlyadvanced to the common carotid artery and then into the internal carotidartery so as to occlude the origin of the middle cerebral artery. Thefilament was withdrawn one hour later to allow reperfusion.

2—Measurement of Brain Infarct Volume:

Twenty-four hours after reperfusion, animals previously treated or notwith the inventive compounds were euthanized by pentobarbital overdose.

Brains were rapidly frozen and sliced. Sections were stained with cresylviolet. Unstained zones of the brain sections were considered to bedamaged by the infarct. Areas (of the infarct and the two hemispheres)were measured and the volume of the infarct and the two hemispheres wascalculated and the corrected infarct volume was determined by thefollowing formula: (corrected infarct volume=infarct volume—(volume ofright hemisphere—volume of left hemisphere)) to compensate for cerebraloedema.

Analysis of the brain sections from animals treated with the inventivecompounds revealed a marked decrease in infarct volume as compared withuntreated animals. When the inventive compounds were administered to theanimals before the ischemia (prophylactic effect), they were capable ofinducing neuroprotection.

3—Measurement of Antioxidant Enzyme Activity:

The rat brains were frozen, crushed and reduced to powder, thenresuspended in saline solution. The different enzyme activities werethen measured as described by the following authors: superoxidedismutase (Flohe and Otting 1984); glutathione peroxidase (Paglia andValentine 1967); glutathione reductase (Spooner, Delides et al. 1981);glutathione-S-transferase (Habig and Jakoby 1981); catalase (Aebi 1984).

Said different enzyme activities were increased in brain preparationsfrom animals treated with the inventive compounds.

B—Curative or Acute Phase Treatment Model:

1—Ischemia Induction/Reperfusion by Intraluminal Occlusion of the MiddleCerebral Artery:

Animals such as those described previously were used for thisexperiment. Animals were anesthetized by intraperitoneal injection of300 mg/kg chloral hydrate. A rectal probe was inserted and bodytemperature was maintained at 37° C.±0.5° C. Blood pressure wasmonitored throughout the experiment. Under a surgical microscope, theright carotid artery was exposed by a median incision in the neck. Thepterygopalatine artery was ligated at its origin and an arteriotomy wasfashioned in the external carotid artery so as to insert a nylonmonofilament, which was gently advanced to the common carotid artery andthen into the internal carotid artery so as to occlude the origin of themiddle cerebral artery. The filament was withdrawn one hour later toallow reperfusion.

2—Treatment of Animals:

Animals first subjected to ischemia-reperfusion were treated with theinventive compounds by the oral route (such as previously described inCMC+Tween vehicle) one or more times after reperfusion (600 mg/kg/day or300 mg/kg/day bid).

3—Measurement of Brain Infarct Volume:

24, 48 or 72 hours after reperfusion, animals previously treated or notwith the compounds were euthanized by pentobarbital overdose.

Brains were rapidly frozen and sliced. Sections were stained with cresylviolet. Unstained zones of the brain sections were considered to bedamaged by the infarct. Areas (of the infarct and the two hemispheres)were measured, the volume of the infarct and the two hemispheres wascalculated and the corrected infarct volume was determined by thefollowing formula: (corrected infarct volume=infarct volume−(volume ofright hemisphere−volume of left hemisphere)) to compensate for cerebraloedema.

In the case of curative treatment (treatment of the acute phase),animals treated with the inventive compounds had less brain damage thanuntreated animals. In fact, the infarct volume was smaller when theinventive compounds were administered for 24, 48 or 72 afterischemia-reperfusion.

The inventive compounds therefore exhibit neuroprotective activityduring treatment following acute ischemia.

The use of the inventive compounds in different experimental modelsshows that said novel compounds have intrinsic antioxidant activity, arecapable of delaying and reducing the effects of an oxidative stress, andfurthermore also induce the expression of genes coding for antioxidantenzymes, which together with their antioxidant property reinforces theprotection against free radicals. In addition, the inventive compoundsexhibit anti-inflammatory activity and are capable of activating thePPARα nuclear receptor

Finally, use of the inventive compounds in an animalischemia-reperfusion model revealed the beneficial neuroprotectiveeffect of both preventive and curative treatment

BIBLIOGRAPHIC REFERENCES

-   Adams, E. P., F. P. Doyle, et al. (1960). “Antituberculous sulphur    compounds. Part IV. Some dimercaptopropyl esters and related    dithiouronium bromides.” J Chem Soc: 2674-80.-   Adams, H. P., Jr. (2002). “Emergent use of anticoagulation for    treatment of patients with ischemic stroke.” Stroke 33(3): 856-61.-   Aebi, H. (1984). “Catalase in vitro.” Methods Enzymol 105: 121-6.-   Antoniadou-Vyzas, A., G. B. Foscolos, et al. (1986). “Di-adamantane    derivatives of a,o-polymethylenediamines with antimicrobial    activity.” Eur J Med Chem Chim Ther 2(1): 73-74.-   Bhatia, S. K. and J. Hajdu (1987). “Stereospecific synthesis of    2-thiophosphatidylcholines; a new class of biologically active    phospholipid analogues.” Tetrahedron Lett 28(33): 3767-3770.-   Bordet, R., D. Deplanque, et al. (2000). “Increase in endogenous    brain superoxide dismutase as a potential mechanism of    lipopolysaccharide-induced brain ischemic tolerance.” J Cereb Blood    Flow Metab 20(8): 1190-6.-   Chang, R. C., P. Hudson, et al. (2000). “Influence of neurons on    lipopolysaccharide-stimulated production of nitric oxide and tumor    necrosis factor-alpha by cultured glia.” Brain Res 853(2): 236-44.-   Clark, R. B. (2002). “The role of PPARs in inflammation and    immunity.” J Leukoc Biol 71(3): 388-400.-   Dimagl, U., C. ladecola, et al. (1999). “Pathobiology of ischaemic    stroke: an integrated view.” Trends Neurosci 22(9): 391-7.-   Farinelli, S. E., D. S. Park, et al. (1996). “Nitric oxide delays    the death of trophic factor-deprived PC12 cells and sympathetic    neurons by a cGMP-mediated mechanism.” J Neurosci 16(7): 2325-34.-   Flohe, L. and F. Otting (1984). “Superoxide dismutase assays.”    Methods Enzmmol 105: 93-104.-   Fruchart, J. C., B. Staels, et al. (2001). “PPARS, metabolic disease    and atherosclerosis.” Pharmacol Res 44(5): 345-   Gaffney, P. R. J. and C. B. Reese (1997). “Preparation of    2-O-arachidonoyl-1-O-stearoyl-sn-glycerol and other di-O-acyl    glycerol derivatives.” Tetrahedron Lett 38(14): 2539-2542.-   Gilgun-Sherki, Y., E. Melamed, et al. (2001). “Oxidative stress    induced-neurodegenerative diseases: the need for antioxidants that    penetrate the blood brain barrier.” Neuropharmacology 40(8): 959-75.-   Gorelick, P. B. (2002). “Stroke prevention therapy beyond    antithrombotics: unifying mechanisms in ischemic stroke pathogenesis    and implications for therapy: an invited review.” Stroke 33(3):    862-75.-   Greene, L. A. and A. S. Tischler (1976). “Establishment of a    noradrenergic clonal line of rat adrenal pheochromocytoma cells    which respond to nerve growth factor.” Proc Natl Acad Sci USA 73(7):    2424-8.-   Gronowitz, S., B. Herslbf, et al. (1978). “Syntheses and chroptical    properties of some derivatives of 1-thioglycerol.” Chem Phys Lipids    22: 307-320.-   Habig, W. H. and W. B. Jakoby (1981). “Assays for differentiation of    glutathione S-transferases.” Methods Enzymol 77: 398-405.-   Jurgens, G., H. F. Hoff, et al. (1987). “Modification of human serum    low density lipoprotein by oxidation—characterization and    pathophysiological implications.” Chem Phys Lipids 45(2-4): 315-36.-   Kainu, T., A. C. Wikstrom, et al. (1994). “Localization of the    peroxisome proliferator-activated receptor in the brain.”    Neuroreport 5(18): 2481-5.-   Kitchin, J., R. C. Bethell, et al. (1994). “Synthesis and    structure-activity relationships of a series of penicillin-derived    HIV proteinase inhibitors: heterocyclic ring systems containing P1′    and P2′ substituents.” J Med Chem 37(22): 3707-16.-   Kotsovolou, S., A. Chiou, et al. (2001). “Bis-2-oxo amide    triacylglycerol analogues: a novel class of potent human gastric    lipase inhibitors.” J Org Chem 66(3): 962-7.-   Lebeau, J., C. Furman, et al. (2000). “Antioxidant properties of    di-tert-butylhydroxylated flavonoids.” Free Radic Biol Med 29(9):    900-12.-   Lutsep, H. L. and W. M. Clark (2001). “Current status of    neuroprotective agents in the treatment of acute ischemic stroke.”    Curr Neurol Neurosci Rep 1(1): 13-8.-   Marx, M. H., C. Piantadosi, et al. (1988). “Synthesis and evaluation    of neoplastic cell growth inhibition of 1-N-alkylamide analogues of    glycero-3-phosphocholine.” J Med Chem 31(4): 858-63.-   Mates, J. M., C. Perez-Gomez, et al. (1999). “Antioxidant enzymes    and human diseases.” Clin Biochem 32(8): 595-603.-   Morliere, P., A. Moysan, et al. (1991). “UVA-induced lipid    peroxidation in cultured human fibroblasts.” Biochim Biophys Acta    1084(3): 261-8.-   Morris, A. D., G. Atassi, et al. (1997). “The synthesis of novel    melphalan derivatives as potential antineoplastic agents.” Eur J Med    Chem 32(4): 343-50.-   Murata, M., S. Ikoma, et al. (1991). “New synthesis of 2-thio-PAF    and related compounds as substrates of PAF acetylhydrolase and    phospholipase A2.” Chem Pharm Bull 39(5): 1335-1336.-   Nandagopal, K., T. M. Dawson, et al. (2001). “Critical role for    nitric oxide signaling in cardiac and neuronal ischemic    preconditioning and tolerance.” J Pharmacol Exp Ther 297(2): 474-8.-   Nazih, A., Y. Cordier, et al. (1999). “Synthesis and stability study    of the new pentaammonio lipidpcTG90, a gene transfer agent.”    Tetrahedron Lett 40(46): 8089-92.-   Nazih, A., Y. Cordier, et al. (2000). “One-pot transformation of a    t-butyl carbamate to a bromoacetamide in the synthesis of the gene    transfer agent pcTG201.” Synlett 5: 635-6.-   Paglia, D. E. and W. N. Valentine (1967). “Studies on the    quantitative and qualitative characterization of erythrocyte    glutathione peroxidase.” J Lab Clin Med 70(1): 158-69.-   Rahman, M. D., D. L. Ziering, et al. (1988). “Effects of    sulfur-containing analogues of stearic acid on growth and fatty acid    biosynthesis in the protozoan Crithidia fasciculata.” J Med Chem    31(8): 1656-9.-   Ramalingan, K., N. Raju, et al. (1995). “Synthesis of nitroimidazole    substituted 3,3,9,9-tetramethyl-4,8-diaza-undecane-2,10-dione    dioximes (propylene amine oximes, PnAOs): ligands for technetium-99m    complexes with potential for imaging hypoxic tissue.” Tetrahedron    51(10): 2875-94.-   Raspe, E., L. Madsen, et al. (1999). “Modulation of rat liver    apolipoprotein gene expression and serum lipid levels by    tetradecylthioacetic acid (TTA) via PPARalpha activation.” J Lipid    Res 40(11): 2099-110.-   Rothwell, N.J. (1997). “Cytokines and acute neurodegeneration.” Mol    Psychiatry 2(2): 120-1.-   Shealy, Y. F., J. L. Frye, et al. (1984). “Synthesis and properties    of some 13-cis- and all-trans-retinamides.” J Pharm Sci 73(6):    745-51.-   Smith, K. J., E. Dipreta, et al. (2001). “Peroxisomes in    dermatology. Part II.” J Cutan Med Surg 5(4): 315-22.-   Spooner, P. J., S. B. Clark, et al. (1988). “The ionization and    distribution behavior of oleic acid in chylomicrons and    chylomicron-like emulsion particles and the influence of serum    albumin.” J Biol Chem 263(3): 1444-53.-   Spooner, R. J., A. Delides, et al. (1981). “Heat stability and    kinetic properties of human serum glutathione reductase activity in    various disease states.” Biochem Med 26(2): 239-48.-   Urakami, C. and K. Kakeda (1953). “Derivatives of    dl-aminopropanediols.” Bull Chem Soc Jpn 26(5): 276-278.

1-17. (canceled)
 18. A compound represented by general formula (I)

in which: G2 and G3 independently represent an oxygen atom, a sulfuratom or a N—R4 group, wherein G2 and G3 do not simultaneously representa N—R4 group, R and R4 independently represent a hydrogen atom or alinear or branched alkyl group, saturated or not, optionallysubstituted, containing from 1 to 5 carbon atoms, R1, R2 and R3, whichare the same or different, represent a hydrogen atom, a CO—R5 group or agroup corresponding to the formula CO—(CH₂)_(2n+1)—X—R6, wherein atleast one of the groups R1, R2 or R3 is a group corresponding to theformula CO—(CH₂)_(2n+1)—X—R6, R5 is a linear or branched alkyl group,saturated or not, optionally substituted, optionally comprising a cyclicgroup, the main chain of which contains from 1 to 25 carbon atoms, X isa sulfur atom, a selenium atom, a SO group or a SO₂ group n is a wholenumber comprised between 0 and 11, R6 is a linear or branched alkylgroup, saturated or not, optionally substituted, optionally comprising acyclic group, the main chain of which contains from 3 to 23 carbonatoms, preferably 10 to 23 carbon atoms and possibly one or moreheterogroups selected in the group consisting of an oxygen atom, asulfur atom, a selenium atom, a SO group and a SO₂ group, with theexception of a compound represented by formula (I) in which G2R2 andG3R3 simultaneously represent hydroxyl groups, the optical andgeometrical isomer, racemate, salt, hydrate thereof and mixturesthereof.
 19. The compound according to claim 18, wherein a single one ofthe groups R1, R2 or R3 represents a hydrogen atom.
 20. The compoundaccording to claim 18, wherein, in the CO—(CH₂)_(2n+1)—X—R6 group, Xrepresents a sulfur or selenium atom and advantageously a sulfur atom.21. The compound according to claim 18, wherein, in theCO—(CH₂)_(2n+1)—X—R6 group, n is comprised between 0 and 3, morespecifically comprised between 0 and 2 and in particular is equal to 0.22. The compound according to claim 18, wherein R6 contains one or moreheterogroups, preferably 0, 1 or 2, more preferably 0 or 1, selected inthe group consisting of an oxygen atom, a sulfur atom, a selenium atom,a SO group and a SO₂ group.
 23. The compound according to claim 18,wherein CO—(CH₂)_(2n+1)—X—R6 is the CO—CH₂—S—C₁₄H₂₉ group.
 24. Thecompound according to claim 18, wherein at least one of the groups R1,R2 and R3 represents a CO—(CH₂)_(2n+1)—X—R6 group in which X representsa sulfur or selenium atom and preferably a sulfur atom and/or R6 is asaturated and linear alkyl group containing from 3 to 23 carbon atoms,preferably 13 to 20 carbon atoms, preferably 14 to 17, more preferably14 to 16, and even more preferably 14 carbon atoms.
 25. The compoundaccording to claim 18, wherein at least two of the groups R1, R2 and R3are CO—(CH₂)_(2n+1)—X—R6 groups, which are the same or different, inwhich X represents a sulfur or selenium atom, preferably a sulfur atom.26. The compound according to claim 18, wherein G2 represents an oxygenor sulfur atom, preferably an oxygen atom.
 27. The compound according toclaim 18, wherein G2 represents an oxygen or sulfur atom and R2represents a group corresponding to the formula CO—(CH₂)_(2n+1)—X—R6.28. The compound according to claim 18, wherein G3 is a N—R4 group inwhich R4 is a hydrogen atom or a methyl group, and G2 is an oxygen atom;and/or R2 represents a CO—(CH₂)_(2n+1)—X—R6 group.
 29. The compoundaccording to claim 18, wherein R1, R2 and R3, which are the same ordifferent, preferably the same, represent a CO—(CH₂)_(2n+1)—X—R6 group,in which X represents a sulfur or selenium atom and preferably a sulfuratom and/or R6 is a saturated and linear alkyl group containing from 13to 17 carbon atoms, preferably 14 to 17, even more preferably 14 carbonatoms, in which n is preferably comprised between 0 and 3, and inparticular is equal to 0, more specifically, R1, R2 and R3 representingCO—CH₂—S—C₁₄H₂₉ groups.
 30. The compound according to claim 18, selectedin the group consisting of:1-tetradecylthioacetylamino-2,3-(dipalmitoyloxy)propane;3-tetradecylthioacetylamino-1,2-(ditetradecylthioacetyloxy)propane;3-palmitoylamino-1,2-(ditetradecylthioacetyloxy)propane;1,3-di(tetradecylthioacetylamino)propan-2-ol;1,3-diamino-2-(tetradecylthioacetyloxy) propane;1,3-ditetradecylthioacetylamino-2-(tetradecylthioacetyloxy)propane;1,3-dioleoylamino-2-(tetradecylthioacetyloxy)propane;1,3-ditetradecylthioacetylamino-2-(tetradecylthioacetythio)propane; and1-tetradecylthioacetylamino-2,3-di(tetradecylthioacetylthio)propane. 31.A pharmaceutical composition comprising, in a pharmaceuticallyacceptable support, at least one compound represented by formula (I)such as defined in any of the previous claims, including a compoundrepresented by formula (I) in which the groups G2R2 and G3R3simultaneously represent hydroxyl groups.
 32. The pharmaceuticalcomposition according to claim 31, for the treatment or prophylaxis ofcerebrovascular pathologies and more particularly cerebral ischemia orstroke.
 33. A method for the treatment of cerebrovascular pathologiesand more particularly cerebral ischemia or stroke, by administering to asubject in need of such treatment an effective amount of a compoundrepresented by formula (I) as defined in claim 18, including a compoundrepresented by formula (I) in which the groups G2R2 and G3R3simultaneously represent hydroxyl groups.